Lost in Translation : Translation Mechanisms in Production of Cocksfoot Mottle Virus Proteins

The present study focuses on the translational strategies of Cocksfoot mottle virus (CfMV, genus Sobemovirus), which infects monocotyledonous plants. CfMV RNA lacks the 5'cap and the 3'poly(A) tail that ensure efficient translation of cellular messenger RNAs (mRNAs). Instead, CfMV RNA is covalently linked to a viral protein VPg (viral protein, genome-linked). This indicates that the viral untranslated regions (UTRs) must functionally compensate for the lack of the cap and poly(A) tail. We examined the efficacy of translation initiation in CfMV by comparing it to well-studied viral translational enhancers. Although insertion of the CfMV 5'UTR (CfMVe) into plant expression vectors improved gene expression in barley more than the other translational enhancers examined, studies at the RNA level showed that CfMVe alone or in combination with the CfMV 3'UTR did not provide the RNAs translational advantage. Mutation analysis revealed that translation initiation from CfMVe involved scanning. Interestingly, CfMVe also promoted translation initiation from an intercistronic position of dicistronic mRNAs in vitro. Furthermore, internal initiation occurred with similar efficacy in translation lysates that had reduced concentrations of eukaryotic initiation factor (eIF) 4E, suggesting that initiation was independent of the eIF4E. In contrast, reduced translation in the eIF4G-depleted lysates indicated that translation from internally positioned CfMVe was eIF4G-dependent. After successful translation initiation, leaky scanning brings the ribosomes to the second open reading frame (ORF). The CfMV polyprotein is produced from this and the following overlapping ORF via programmed -1 ribosomal frameshift (-1 PRF). Two signals in the mRNA at the beginning of the overlap program approximately every fifth ribosome to slip one nucleotide backwards and continue translation in the new -1 frame. This leads to the production of C-terminally extended polyprotein, which encodes the viral RNA-dependent RNA polymerase (RdRp). The -1 PRF event in CfMV was very efficient, even though it was programmed by a simple stem-loop structure instead of a pseudoknot, which is usually required for high -1 PRF frequencies. Interestingly, regions surrounding the -1 PRF signals improved the -1 PRF frequencies. Viral protein P27 inhibited the -1 PRF event in vivo, putatively by binding to the -1 PRF site. This suggested that P27 could regulate the occurrence of -1 PRF. Initiation of viral replication requires that viral proteins are released from the polyprotein. This is catalyzed by viral serine protease, which is also encoded from the polyprotein. N-terminal amino acid sequencing of CfMV VPg revealed that the junction of the protease and VPg was cleaved between glutamate (E) and asparagine (N) residues. This suggested that the processing sites used in CfMV differ from the glutamate and serine (S) or threonine (T) sites utilized in other sobemoviruses. However, further analysis revealed that the E/S and E/T sites may be used to cleave out some of the CfMV proteins.

Abductive translation studies : The art of marshalling signs

This dissertation contributes to the fields of theoretical translation studies, semiotic translation theory, and the semiotics of translation. The aim of this work is to explore the alternative and potential which the semiotic approaches to translation entail from the viewpoint of contemporary translation studies. The overall objective is thus to show that a general semiotic translation theory, and in particular, a Peircean translation theory, are possible and indispensable. Furthermore, this study contributes to the semiotranslational approach and to its theory-building by developing the concept of abductive translation (studies). The specific theoretical frame of reference adopted in this study is provided by the semiotranslation introduced by Dinda L. Gorlée. This approach is primarily based on the semeiotic of Charles Sanders Peirce (1839 1914), and aims at a fusion of semiotics and translation studies. A more general framework is provided by the threefold background and material: the published and unpublished writings of Peirce, Peirce scholarship and Peircean-semiotic publications, as well as the translation-theoretical literature. Part One of this study concentrates on the justification, existence, and nature of the semiotic approaches to translation. This part provides a historical survey, a status report, and a discussion of this area of research, by employing the findings in a boundary-clearing that is multilayered both conceptually and terminologically. Part Two deals with Peircean semiotranslation. Here Gorlée s semiotranslational research is examined by focusing on the starting points, features, and development of semiotranslation. Attention is also paid to the state-of-the-art of semiotranslation theory and to the possibilities for future elaborations. Part Three focuses on the semiotranslational claim that translation is an abductive activity. The concept of abductive translation is based on abduction, one of Peirce s three modes of reasoning; at the same time Firstness, the category of abduction, becomes foregrounded. So abductive translation as a form of possibilistic translation receives here an extensive theoretical discussion by citing examples in which abduction manifests itself as (scientific) reasoning and as everyday contemplation. During this treatise, translation is first equated with sign action, then with interpretation and finally with reasoning. All these approaches appear to embody different facets of the same phenomenon Peirce s ubiquitous semiosis, and they all suggest that translation is inherently an intersemiotic activity in which a sign is inferred from another sign. Translation is therefore semiosis, semiosis is translation and interpretation, interpretation is reasoning, and so on ad infinitum all being manifestations of the art of marshalling signs. The three parts of this study are linked by the overall goal of abductive translation studies: investigation into abductive translation develops the theory of semiotranslation, and this enrichment of semiotranslation in turn constructs a semiotic paradigm within translation studies.

The Duet between the Author and the Translator: An Analysis of Style through Shifts in Literary Translation

The study proposes a method for identifying the personal imprint of literary translators in translated works of fiction. The initial assumption was that the style of a target text is not determined solely by the literary style of the author but also by features of its translator s idiolect. A method was developed for identifying the idiolectal features of individual translators, which were then used to describe personal translation styles. The method is not restricted to a particular language pair. To test the method and to establish the nature of the proposed personal imprint empirically, extracts from four English-language literary source texts (two novels by James Joyce and two by Ernest Hemingway) were first compared with their translations into Finnish (by four different translators) in order to identify changes, or shifts, that had taken place at the formal linguistic level in the translation process. To allow individual propensities to manifest themselves, only optional shifts in which the translators had a range of choices available to them were included in the study. In the second phase, extracts by different authors rendered into Finnish by the same translator were compared in order to gauge the extent of the potential impact of the author's style on the translator's work. In-depth analysis of the types of shifts made most frequently by the individual translators revealed further intersubjective differences, and the shifts were used to construct translation profiles for each of the translators. In order to determine the potential effects of frequently occurring shifts on the target text, some central concepts of narratology were adapted and used to establish an intermediate link between microlevel choices and macrolevel effects. In this way the propensity of an individual translator to opt for certain types of shift could be linked with the overall artistic effect of the target text.

Measuring attitudes in translation: a study of Nokia business reports

Background Contemporary Finnish, spoken and written, reveals loanwords or foreignisms in the form of hybrids: a mixture of Finnish and foreign syllables (alumiinivalua). Sometimes loanwords are inserted into the Finnish sentence in their raw form just as they are found in the source language (pulp, after sales palvelu). Again, sometimes loanwords are calques, which appear Finnish but are spelled and pronounced in an altogether foreign manner (Protomanageri, Promenadi kampuksella). Research Questions What role does Finnish business translation play in the migration of foreignisms into Finnish if we consider translation "as a construct of solutions determined by the ideological constraints and conflicts characterizing the target culture" (Robyns 1992: 212)? What attitudes do the Finns display toward the presence of foreignisms in their language? What socio-economic or ideological conditions (Bassnett 1994: 321) are responsible for these attitudes? Are these conditions dynamic? What tools can be used to measure such attitudes? This dissertation set out to answer these and similar questions. Attitudes are imperialist (where otherness is both denied and transformed), defensive (where otherness is acknowledged, transformed, and vilified), transdiscursive (a neutral attitude to both otherness and transformation), or finally defective (where alien migration is acknowledged and "stimulated") (Robyns 1994: 60). Methodology The research method follows Rose's schema (1984: 8): (a) take an existing theory, (b) develop from it a proposition specific enough to be tested, (c) devise a scheme that tests this proposition, (d) carry through the scheme in practice, (e) draw up results and discuss conclusions in relation to the original theory. In other words, the method attempts an explanation of a Finnish social phenomenon based on systematic analyses of translated evidence (Lewins 1992: 4) whereby what really matters is the logical sequence that connects the empirical data to the initial research questions raised above and, ultimately to its conclusion (Yin 1984: 29). Results This research found that Finnish translators of the Nokia annual reports used a foreignism whenever possible such as komponentin instead of rakenneosa, or investoida instead of sijoittaa, and often without any apparent justification (Pryce 2003: 203-12) more than the translator's personal preference. In the old documents (minutes of meetings of the Board of Directors of Osakeyhtio H. Saastamoinen, Ltd. dated 5 July 1912-1917, a NOPSA booklet (1932), Enzo-Gutzeit-Tornator Oy document (1938), Imatra Steel Oy Annual Report 1964, and Nokia Oy Annual Report 1946), foreignisms under Haugen's (1950: 210-31) Classification #1 occurred an average of 0.6 times, while in the new documents (Nokia 1998 translated Annual Reports) they occurred an average of 6.5 times. That big difference, suggests transdiscursive and defective attitudes in Finnish society toward the other. In the 1850s, Finnish attitudes toward alien persons and cultures were hardened, intolerant and prohibitive because language politics were both nascent and emerging, and Finns adopted a defensive stance (Paloposki 2002: 102 ff) to protect their cultural and national treasures such as language and folklore. Innovation The innovation here is that no prior doctoral level research measured Finnish attitudes toward foreignisms using a business translation approach. This is the first time that Haugen's classification has been modified and applied in target language analysis. It is hoped that this method would be replicated in similar research in the future. Applications For practical applications, researchers with interest in languages, language development, language influences, language ideologies, and power structures that affect national language policies will find this thesis useful, especially the model for collecting, grouping, and analyzing foreignisms that has been demonstrated here. It is intended to document for posterity current attitudes of Finns toward the other as revealed in business translations from 1912-1964, and in 1998. This way, future language researchers would be able to explore a time-line of Finnish language development and attitudes toward the other. Communication firms may also find this research interesting. In future, could the model we adopted be used to analyze literary texts or religious texts for example? Future Trends Though business documents show transdiscursive attitudes, other segments of Finnish society may show defensive or imperialist attitudes. When the ideology of industrialization changes in the future, will Finnish attitudes toward the other change as well? Will it then be possible to use the same kind of analytical tools to measure Finnish attitudes? More broadly, will linguistic change continue in the same direction of transdiscursive attitudes, or will the change slow down or even reverse into xenophobic attitudes? Is this our model culture-specific or can it be used in the context of other cultures? Conclusion There is anger against foreignisms in Finland as newspaper publications and television broadcasts show, but research shows that a majority of Finns consider foreignisms and the languages from which they come as sources of enrichment for Finnish culture (Laitinen 2000, Eurobarometer series 41 of July 1994, 44 of Spring 1996, 50 of Autumn 1998). Ideologies of industrialization and globalization in Finland have facilitated transdiscursive tendencies. When Finland's political ideology was intolerant toward foreign influences in the 1850s because Finland was in the process of consolidating her nascent country and language, attitudes toward the importation of loanwords also became intolerant. Presently, when industrialization and globalization became the dominant ideologies, we see a shift in attitudes toward transdiscursive tendencies. Ideology is usually unseen and too often ignored by translation researchers. However, ideology reveals itself as the most powerful factor affecting language attitudes in a target culture. Key words Finnish, Business Translation, Ideology, Foreignisms, Imperialist Attitudes, Defensive Attitudes, Transdiscursive Attitudes, Defective Attitudes, the Other, Old Documents, New Documents.

Realia, Translation Strategies, and Subtitling : A study of two comedy sketch shows on DVD

Tutkielman tavoitteena on tutkia, millaisia käännösstrategioita DVD-tekstittämisessä suositaan reaalioita eli kulttuurisidonnaisia käsitteitä käännettäessä. Lähtökohtana on, että osaa strategioista voidaan käyttää reaalioiden kulttuurisen erilaisuuden korostamiseen, osaa taas tämän erilaisuuden häivyttämiseen. Nämä kaksi kategoriaa eivät kuitenkaan kata kaikkia mahdollisia strategioita, minkä vuoksi myös neutraalit strategiat otetaan tutkielmassa huomioon. Tutkielman aineisto koostuu Suomessa nimillä Ruuvit löysällä ja Pikku-Britannia tunnettujen brittiläisten sketsisarjojen DVD-julkaisuista. Molemmat tutkituista julkaisuista käsittävät yhden tuotantokauden. Formaatiltaan sketsisarjat perustuvat lukuisiin lyhyisiin kohtauksiin, joissa käsitellään useita eri aihealueita. Tämän vuoksi ne soveltuvat erinomaisesti reaalioiden tutkimukseen. Tutkittava kääntämisen laji on DVD-kääntäminen, koska sen merkityksen voidaan katsoa kasvaneen DVD:n lisääntyneen suosion myötä. Tutkielman teoriatausta muodostuu reaalioiden, käännösstrategioiden ja tekstityksen teoriasta. Näistä osa-alueista ensimmäisessä korostuu kulttuurin vaikutus kieleen ja siten myös kääntämiseen, kun taas toinen pohjautuu sekä tekstitason kotouttavista ja vieraannuttavista käännösstrategioista että sanatason lokaaleista strategioista esitettyyn teoriaan, ja kolmannessa nousevat esille sanan ja kuvan yhteistyö sekä muut tekstittämiseen vaikuttavat tekijät, kuten rajallinen käytettävissä oleva tila ja aika. Yhdessä teorian eri osa-alueet mahdollistavat tutkimuksen, jossa voidaan tarkastella kulttuuristen, kielellisten ja audiovisuaalisten piirteiden vaikutusta reaalioiden kääntämiseen. Tutkielmassa käytetään kvalitatiivista ja kvantitatiivista tutkimusmenetelmää. Kvalitatiivinen menetelmä perustuu käännösstrategioiden käyttötapojen kuvaamiseen tutkimalla tutkimusaineistosta transkriboituja reaalioiden käännöksiä yksittäisissä tapauksissa. Analyysissa otetaan huomioon ensisijaisesti reaalioiden tekstuaalinen ja audiovisuaalinen konteksti sekä niiden oletettu tunnistettavuus käännösten kohdekulttuurissa. Tätä analyysia täydennetään kaikkien havaittujen reaalioiden kääntämistä kuvaavalla kvantitatiivisella menetelmällä. Tutkielman tulokset viittaavat siihen, että erilaisten käännösstrategioiden välillä on huomattavia eroja siinä, millä tavoin ja miten usein niitä käytetään. Lisäksi tulokset osoittavat odotetusti sen, että kotouttavat ja vieraannuttavat strategiat eivät sulje toisiaan pois, vaan niitä molempia voidaan käyttää saman käännöksen sisällä. Kokonaisuutena tutkielmassa tuodaan ilmi monia aihealueen tutkimiseen liittyviä haasteita aina kulttuurin määrittelystä audiovisuaalisen aineiston analysoimiseen asti. Avainsanat: Kulttuurisidonnaiset käsitteet, käännösstrategia, tekstittäminen, kotouttaminen, vieraannuttaminen

Crime stories from the country of huopatossu and juhannusruusut : Translation strategies for Finnish cultural realia in the English translations of two of Matti Yrjänä Joensuu’s crime novels

Pro gradu –tutkielman aiheena on kulttuurin ominaispiirteiden kääntäminen. Teksteissä kulttuurin jälki voi näkyä monin eri tavoin. Tutkielman kohteena ovat erityisesti käännösstrategiat, joita kääntäjät käyttävät kohdatessaan kulttuurisidonnaisia viittauksia. Tutkielma nostaa esiin myös niitä tekijöitä, jotka vaikuttavat siihen, minkälaisia käännösstrategioita kääntäjät valitsevat. Näistä tekijöistä tutkielma keskittyy kääntämisen normeihin. Tutkielma pureutuu kulttuurisidonnaisten viitteiden kääntämiseen tarkastelemalla kahta suomalaista kaunokirjallista teosta ja niiden käännöksiä. Tutkielman aineistona ovat Matti Yrjänä Joensuun kaksi rikosromaania ja näiden käännökset. Teoksista toinen on vuonna 1983 suomeksi julkaistu Harjunpää ja poliisin poika, jonka englanninkielinen käännös Harjunpaa the stone murders julkaistiin vuonna 1986. Toinen teos on vuonna 2003 julkaistu Harjunpää ja pahan pappi ja sen käännös The Priest of Evil vuodelta 2006. Tutkielman tavoitteena on selvittää, minkälaisia kulttuurisidonnaisia viittauksia romaanit sisälsivät, minkälaisia käännösstrategioita kääntäjät käyttivät kääntäessään näitä viittauksia ja mikä voisi selittää heidän strategisia valintojaan. Tutkielma pyrkii vastaamaan kysymykseen siitä, voisiko jonkin kääntämistä koskevan normin olemassa olo selittää kääntäjien strategisia valintoja. Tutkielman tavoitteena on myös selvittää, suositaanko kääntämisessä englannin kieleen niin kutsuttuja kotouttavia käännösstrategioita ja ovatko käännösstrategiat ja kääntämistä koskevat normit muuttuneet kahdenkymmenen vuoden aikana. Näihin kysymyksiin vastaamiseksi suomenkielisistä teksteistä on etsitty kaikki kulttuurisidonnaisia viittauksia sisältävät tekstinkohdat. Näitä vastaavat kohdat on sitten etsitty käännöksistä ja suomen- ja englanninkielisiä kohtia on vertailtu keskenään. Molemmat suomenkieliset romaanit sisältävät runsaasti kulttuurisidonnaisia viittauksia. Suurimman kulttuurisidonnaisten viittausten ryhmän muodostivat molemissa romaaneissa henkilöiden nimet. Romaanin sisälsivät myös runsaasti viittauksia maantieteeseen, erityisesti kulttuurimaantieteeseen, ja yhteiskuntaan. Sitä vastoin viittaukset suomalaiseen kulttuuriin ja historiaan olivat vähäisempiä. Tutkielma osoittaa, että kääntäessään suomesta englannin kielelle suomalaisen rikoskirjallisuuden kääntäjät saattavat käyttää enemmän vieraannuttavia strategioita kuin kotouttavia strategioita ja että he suosivat vieraannuttavia strategioita kasvavassa määrin. Harjunpään ja pahan papin kääntäjä käytti enemmän vieraannuttavia strategioita kuin Harjunpään ja poliisin pojan kääntäjä kaksikymmentä vuotta aikaisemmin. Tutkielman tulokset eivät tue väitettä siitä, että käännettäessä englannin kielelle suosittaisiin kotouttavia strategioita. Näyttää siltä, että vieraannuttavia strategioita on käytetty enemmän ja käytetään yhä enenevässä määrin. Lisääntyvän vieraannuttamisen taustalla voi olla useita syitä, kuten suomalaisen kulttuurin lisääntynyt tunnettuus maailmalla, rikosromaanin genren vaatimukset tai muutokset kääntäjäyhteisön arvoissa. Tutkielman tulosten perusteella näyttää siltä, että ainakin muutoksia normeissa ja arvoissa on tapahtunut. Lisätutkimuksen avulla voitaisiin selvittää, pätevätkö tutkielman tulokset muihin romaaneihin ja niiden käännöksiin tai muihin genreihin käännettäessä suomesta englantiin. Lisätutkimus voisi nojautua laajempaan ja erilaisia tekstejä kattavaan aineistoon. Jatkotutkimus voisi myös sisältää kääntäjien haastatteluita tai kyselyitä kääntäjille. Näiden avulla voitaisiin saada lisäselvyyttä syistä heidän strategisille valinnoilleen. Asiasanat: Käännöskirjallisuus – kaunokirjallisuus Kääntäminen – suomen kieli – englannin kieli Kääntäminen – strategia Kääntäminen - normi

Lost in translation? : Translating allusions in two of Reginald Hill's Dalziel & Pascoe novels

This study examines strategies used to translate various thematic and character delineating allusions in two of Reginald Hill's detective novels, The Wood Beyond and On Beulah Height and their Swedish translations Det mörka arvet and Dalen som dränktes. In this study, thematic allusions and allusions used in character delineation are regarded as intertextual networks. Intertextual networks comprise all the texts that are in one way or another embedded into a text, all the texts referred to in it and even the texts somehow rejected from a text's own canon. Studying allusions as intertextual networks makes it warranted to pay minute attention to even the smallest of details. Seen together, these little details form extensive networks of meaning that readers use to interpret the text. Allusion can be defined as a reference, often covert or indirect, to another text in a way that brings into the text some of the associations of that other text. A text is here understood broadly, hence sources of allusions include all cultural texts from literature and history to cinema and televisions serials. Allusions are culture bound and each culture tends to allude to its own cultural products. The set of transcultural allusions is therefore fairly small. Translation strategies are translatorial ways of solving translation problems. Being culture-bound, allusions are potential translation problems. In order to transmit the thoughts evoked by the allusions in source text readers to the target text readers translators may add guidance to the translated text. Often guidance is not added, which may result in changes in handling of themes or character delineation, clear in the source text but confusing or incomprehensible in the target text. However, norms in target culture may not always allow the translators the possibility to make the text comprehensible. My analyses of translation strategies show that in the two translated novels studied minimum change is a very frequently used strategy. This results in themes and character delineation losing some of the effect they have in the source texts. Perhaps surprisingly, the result is very much the same even where it is possible to discern that the two translators have had differing translation principles. Keywords: allusions, intertextuality, literary translation, translation strategies, norms, crime fiction, Hill, Reginald

From Iron Horse to Rauta-airot - a Semiotic View on the Domestication of Metaphors in Literary Translation

The thesis concentrates on two questions: the translation of metaphors in literary texts, and the use of semiotic models and tools in translation studies. The aim of the thesis is to present a semiotic, text based model designed to ease the translation of metaphors and to analyze translated metaphors. In the translation of metaphors I will concentrate on the central problem of metaphor translations: in addition to its denotation and connotation, a single metaphor may contain numerous culture or genre specific meanings. How can a translator ensure the translation of all meanings relevant to the text as a whole? I will approach the question from two directions. Umberto Eco's holistic text analysis model provides an opportunity to concentrate on the problematic nature of metaphor translation from the level of a text as a specific entity, while George Lakoff's and Mark Johnson's metaphor research makes it possible to approach the question from the level of individual metaphors. On the semiotic side, the model utilizes Eero Tarasti's existential semiotics supported by Algirdas Greimas' actant model and Yuri Lotman's theory of cultural semiotics. In the model introduced in the thesis, individual texts are deconstructed through Eco's model into elements. The textual roles and features of these elements are distilled further through Tarasti's model into their coexistent meaning levels. The priorization and analysis of these meaning levels provide an opportunity to consider the contents and significance of specific metaphors in relation to the needs of the text as a whole. As example texts, I will use Motörhead's hard rock classic Iron Horse/Born to Lose and its translation into Rauta-airot by Viikate. I will use the introduced model to analyze the metaphors in the source and target texts, and to consider the transfer of culture specific elements between the languages and cultural borders. In addition, I will use the analysis process to examine the validity of the model introduced in the thesis.

Translation of Allusions in the Animated Cartoon The Simpsons

Tässä tutkielmassa tarkastellaan The Simpsons (Simpsonit) animaatiosarjassa esiintyvien sanallisten alluusioiden kääntymistä suomenkielisiksi tekstityksiksi. Tarkoituksena on selvittää kvantitatiivisen analyysin keinoin, miten kääntäjä Sari Luhtanen käyttää eri käännösstrategioita hyväkseen. Tutkimusaineisto on kerätty kuudesta Simpsonit-sarjan jaksosta esityskausilta 7-11. Työn teoriaosiossa määritellään aluksi intertekstuaalisuuden sekä alluusion käsitteet, jonka jälkeen käsitellään alluusiota huumorin välineenä. Mahdollisten käännösstrategioiden määrittely tukeutuu Ritva Leppihalmeen alluusioiden kääntämistä käsittelevään tutkimukseen. Leppihalmeen terminologiaa mukaillen alluusiot jaetaan erisnimialluusioihin ja suoriin tekstilainoihin. Simpsonit-sarjan olemusta ja tekijöiden tarkoitusperiä pohtii oma lukunsa, joka sisältää myös kääntäjä Luhtasen ajatuksia käännöstyöstä, sekä haastattelussa esiintulleita yksityiskohtia käännösprosessista. Tekstittämisen erityispiirteitä kääntämisen alalajina käsitellään Henrik Gottliebin sekä Jan Ivarssonin määritelmien kautta. Gottliebin määritelmää elokuvasta polysemioottisena, neljästä eri viestintäkanavasta koostuvana kokonaisuutena sovelletaan myös tutkimuksen empiirisessä osuudessa. Tekstittämiseen todetaan kohdistuvan merkittäviä teknisiä rajoituksia, jotka tehokkaasti estävät kääntäjää pyrkimästä muodolliseen vastaavuuteen lähdetekstin kanssa. Koska tekstitys kuitenkin esitetään aina alkuperäisen materiaalin yhteydessä, sen voi mieltää ylimääräiseksi viestintäkanavaksi jonka avulla kääntäjä auttaa kohdeyleisöä kokemaan tekstitetyn ohjelman alkuperäisen yleisön kokemusta vastaavalla tavalla (dynaaminen vastaavuus). Tutkimuksen empiirinen osuus tarkastelee materiaalissa esiintyviä alluusioiden käännöksiä kvantitatiivisen analyysin muodossa, jonka jälkeen Luhtasen tekemiä käännösvalintoja käsitellään yksityiskohtaisemmin esimerkkien avulla. Luhtasen todetaan sisällyttävän dialogissa esiintyvät alluusiot tekstityksiin lähes aina, mutta useimmiten jättävän tekstityksistä pois pelkästään visuaalisella kanavalla esiintyvät sanallisen alluusiot. Erisnimialluusiot Luhtanen kääntää tyypillisesti muuttumattomina, mutta harkintansa mukaan saattaa Suomessa täysin tuntemattomien nimien kohdalla tarjota myös selittävän käännöksen. Suoria tekstilainoja kääntäessään hän ei näytä suosivan mitään tiettyä strategiaa. Erillisiä selityksiä sisältäviä strategioita Luhtanen ei käytä. Yleisen alluusioiden runsauden lisäksi Simpsonit-sarjan erityispiirteenä näyttävät olevan visuaalisella kanavalla ajoittain esiintyvät, nopeasti ohi menevät sanallisten alluusioiden keskittymät. Näiden alluusiokeskittymien edessä kääntäjä on usein voimaton, ja Luhtanen onkin lähes aina jättänyt tällaiset alluusiokeskittymät kokonaan suomentamatta.

Defects in tricarboxylic acid cycle enzymes Fumarate hydratase and Succinate dehydrogenase in cancer

Hereditary leiomyomatosis and renal cell cancer (HLRCC) is a recently characterized cancer syndrome which predisposes to cutaneous and uterine leiomyomas as well as renal cell carcinoma (RCC). Uterine leiomyosarcoma (ULMS) has also been observed in certain Finnish HLRCC families. The predisposing gene for this syndrome, fumarate hydratase (FH), was identified in 2002. The well-known function of FH is in the tricarboxylic acid cycle (TCAC) in the energy metabolism of cells. As FH is a novel cancer gene, the role of FH mutations in tumours is in general unknown. Similarly, the mechanisms through which defective FH is associated with tumourigenesis are unclear. The loss of a wild type allele has been observed in virtually all HLRCC patients tumours and the FH enzyme activities are either totally lost or remarkably reduced in the tissues of mutation carrier patients. Therefore, FH is assumed to function as a tumour suppressor. Mutations in genes encoding subunits of other TCAC enzyme SDH have also been reported recently in tumours: mutations in SDHB, SDHC, and SDHD genes predispose to paraganglioma and pheochromocytoma. In the present study, mutations in the SDHB gene were observed to predispose to RCC. This was the first time that mutations in SDHB have been detected in extra-paraganglial tumours. Two different SDHB mutations were observed in two unrelated families. In the first family, the index patient was diagnosed with RCC at the age of 24 years. Additionally, his mother with a paraganglioma (PGL) of the heart and his maternal uncle with lung cancer were both carriers of the mutation. The RCC of the index patient and the PGL of his mother showed LOH. In the other family, an SDHB mutation was detected in two siblings who were both diagnosed with RCC at the ages of 24 and 26 years. Both of the siblings also suffered PGL. All these tumours showed LOH. Therefore, we concluded that mutations in SDHB predispose also for RCC in certain families. Several tumour types were analysed for FH mutations to define the role of FH mutations in these tumour types. In addition, patients with a putative cancer phenotype were analysed to identify new HLRCC families. Three FH variants were detected, of which two were novel. One of the variants was observed in a patient diagnosed with ULMS at the age of 41 years. However, LOH was not detected in the tumour tissue. The FH enzyme activity of the mutated protein was clearly reduced, being 43% of the activity of the normal protein. Together with the results from an earlier study we calculated that the prevalence of FH mutations in Finnish non-syndromic ULMS is around 2.4%. Therefore, FH mutations seem to have a minor role in the pathogenesis on non-syndromic ULMS. Two other germline variants were detected in a novel tumour type, ovarian mucinous cystadenoma. However, tumour tissues of the patients were not available for LOH studies and therefore LOH status remained unclear. Therefore, it is possible that FH mutations predispose also for ovarian tumours but further studies are needed to verify this result. A novel variant form of the FH gene (FHv) was identified and characterized in more detail. FHv contains an alternative first exon (1b), which appeared to function as 5 UTR sequence. The translation of FHv is initiated in vitro from exons two and three. The localization of FHv is both cytosolic and nuclear, in contrast to the localization of FH in mitochondria. FHv is expressed at low levels in all human tissues. Interestingly, the expression was induced after heat shock treatment and in chronic hypoxia. Therefore, FHv might have a role e.g. in the adaptation to unfavourable growth conditions. However, this remains to be elucidated.

Sweet potato chlorotic stunt virus: Studies on viral synergism and suppression of RNA silencing

The studies presented in this thesis aimed to a better understanding of the molecular biology of Sweet potato chlorotic stunt virus (SPCSV, Crinivirus, Closteroviridae) and its role in the development of synergistic viral diseases. The emphasis was on the severe sweet potato virus disease (SPVD) that results from a synergistic interaction of SPCSV and Sweet potato feathery mottle virus (SPFMV, Potyvirus, Potyviridae). SPVD is the most important disease affecting sweetpotato. It is manifested as a significant increase in symptom severity and SPFMV titres. This is accompanied by a dramatic sweetpotato yield reduction. SPCSV titres remain little affected in the diseased plants. Viral synergistic interactions have been associated with the suppression of an adaptive general defence mechanism discovered in plants and known as RNA silencing. In the studies of this thesis two novel proteins (RNase3 and p22) identified in the genome of a Ugandan SPCSV isolate were shown to be involved in suppression of RNA silencing. RNase3 displayed a dsRNA-specific endonuclease activity that enhanced the RNA-silencing suppression activity of p22. Comparative analyses of criniviral genomes revealed variability in the gene content at the 3´end of the genomic RNA1. Molecular analyses of different isolates of SPCSV indicated a marked intraspecific heterogeneity in this region where the p22 and RNase3 genes are located. Isolates of the East African strain of SPCSV from Tanzania and Peru and an isolate from Israel were missing a 767-nt fragment that included the p22 gene. However, regardless of the absence of p22, all SPCSV isolates acted synergistically with SPFMV in co-infected sweetpotato, enhanced SPFMV titres and caused SPVD. These results showed that p22 is dispensable for development of SPVD. The role of RNase3 in SPVD was then studied by generating transgenic plants expressing the RNase3 protein. These plants had increased titres of SPFMV (ca. 600-fold higher in comparison with nontransgenic plants) 2-3 weeks after graft inoculation and displayed the characteristic SPVD symptoms. RNA silencing suppression (RSS) activity of RNase3 was detected in agroinfiltrated leaves of Nicotiana bethamiana. In vitro studies showed that RNase3 was able to cleave small interferring RNAs (siRNA) to products of ~14-nt. The data thus identified RNase3 as a suppressor of RNA silencing able to cleave siRNAs. RNase3 expression alone was sufficient for breaking down resistance to SPFMV in sweetpotato and for the development of SPVD. Similar RNase III-like genes exist in animal viruses which points out a novel and possibly more general mechanism of RSS by viruses. A reproducible method of sweetpotato transformation was used to target RNA silencing against the SPCSV polymerase region (RdRp) with an intron-spliced hairpin construct. Hence, engineered resistance to SPCSV was obtained. Ten out of 20 transgenic events challenged with SPCSV alone showed significantly reduced virus titres. This was however not sufficient to prevent SPVD upon coinfection with SPFMV. Immunity to SPCSV seems to be required to control SPVD and targeting of different SPCSV regions need to be assessed in further studies. Based on the identified key role of RNase3 in SPVD the possibility to design constructs that target this gene might prove more efficient in future studies.

Movement-associated proteins of Potato virus A: attachment to virus particles and phosphorylation

The particles of Potato virus A (PVA; genus Potyvirus) are helically constructed filaments that contain multiple copies of a single type of coat-protein (CP) subunit and a single copy of genome-linked protein (VPg), attached to one end of the virion. Examination of negatively-stained virions by electron microscopy revealed flexuous, rod-shaped particles with no obvious terminal structures. It is known that particles of several filamentous plant viruses incorporate additional minor protein components, forming stable complexes that mediate particle disassembly, movement or transmission by insect vectors. The first objective of this work was to study the interaction of PVA movement-associated proteins with virus particles and how these interactions contribute to the morphology and function of the virus particles. Purified particles of PVA were examined by atomic force microscopy (AFM) and immuno-gold electron microscopy. A protrusion was found at one end of some of the potyvirus particles, associated with the 5' end of the viral RNA. The tip contained two virus-encoded proteins, the genome-linked protein (VPg) and the helper-component proteinase (HC-Pro). Both are required for cell-to-cell movement of the virus. Biochemical and electron microscopy studies of purified PVA samples also revealed the presence of another protein required for cell-to-cell movement the cylindrical inclusion protein (CI), which is also an RNA helicase/ATPase. Centrifugation through a 5-40% sucrose gradient separated virus particles with no detectable CI to a fraction that remained in the gradient, from the CI-associated particles that went to the pellet. Both types of particles were infectious. AFM and translation experiments demonstrated that when the viral CI was not present in the sample, PVA virions had a beads-on-a-string phenotype, and RNA within the virus particles was more accessible to translation. The second objective of this work was to study phosphorylation of PVA movement-associated and structural proteins (CP and VPg) in vitro and, if possible, in vivo. PVA virion structural protein CP is necessary for virus cell-to-cell movement. The tobacco protein kinase CK2 was identified as a kinase phosphorylating PVA CP. A major site of CK2 phosphorylation in PVA CP was identified as a single threonine within a CK2 consensus sequence. Amino acid substitutions affecting the CK2 consensus sequence in CP resulted in viruses that were defective in cell-to-cell and long-distance movement. The CK2 regulation of virion assembly and cell-to-cell movement by phosphorylation of CP was possibly due to the inhibition of CP binding to viral RNA. Four putative phosphorylation sites were identified from an in vitro phosphorylated recombinant VPg. All four were mutated and the spread of mutant viruses in two different host plants was studied. Two putative phosphorylation site mutants (Thr45 and Thr49) had phenotypes identical to that of a wild type (WT) virus infection in both Nicotiana benthamiana and N. tabacum plants. The other two mutant viruses (Thr132/Ser133 and Thr168) showed different phenotypes with increased or decreased accumulation rates, respectively, in inoculated and the first two systemically infected leaves of N. benthamiana. The same mutants were occasionally restricted to single cells in N. tabacum plants, suggesting the importance of these amino acids in the PVA infection cycle in N. tabacum.

The Pseudomonas syringae-derived HrpA pilins - molecular characterization and biotechnological application of the transcripts

Many Gram-negative bacteria pathogenic to plants and animals possess type III secretion systems that are used to cause disease. Effector proteins are injected into host cells using the type III secretion machineries. Despite vigorous studies, the nature of the secretion signal for type III secreted proteins still remains elusive. Both mRNA and proteinaceous signals have been proposed. Findings on coupling of translation to secretion by the type III secretion systems are also still contradictory. This study dealt with the secretion signal of HrpA from Pseudomonas syringae pathovar tomato. HrpA is the major component of the type III secretion system-associated Hrp pilus and a substrate for the type III secretion systems. The secretion signal was shown to reside in the first 15 codons or amino acids, a location typical for type III secretion signals. Translation of HrpA in the absence of a functional type III secretion system was established, but it does not exclude the possibility of coupling of translation to secretion when the secretion apparatus is present. The hrpA transcripts from various unrelated plant pathogenic bacteria were shown to be extremely stable. The biological relevance of this observation is unknown, but possible explanations include the high prevalence of HrpA protein, an mRNA secretion signal or timing of secretion. The hrpA mRNAs are stable over a wide range of temperatures, in the absence of translating ribosomes and even in the heterologous host Escherichia coli. The untranslated regions (UTRs) of hrpA transcripts from at least 20 pathovars of Pseudomonas syringae are highly homologous, whilst their coding regions exhibit low similarity. The stable nature of hrpA messenger RNAs is likely to be due to the folding of their 5 and 3 UTRs. In silico the UTRs seem to form stem-loop structures, the hairpin structures in the 3 UTRs being rich in guanidine and cytosine residues. The stable nature of the hrpA transcript redirected the studies to the stabilization of heterologous transcripts and to the use of stable messenger RNAs in recombinant protein production. Fragments of the hrpA transcript can be used to confer stability on heterologous transcripts from several sources of bacterial and eukaryotic origin, and to elevate the levels of production of the corresponding recombinant proteins several folds. hrpA transcript stabilizing elements can be used for improving the yields of recombinant proteins even in Escherichia coli, one of the most commonly used industrial protein production hosts.