Translational coupling by modulation of feedback repression in the IF3 operon of Escherichia coli

A pseudoknot formed by a long-range interaction in the mRNA of the initiation factor 3 (IF3) operon is involved in the translational repression of the gene encoding ribosomal protein L35 by another ribosomal protein, L20. The nucleotides forming the 5′ strand of the key stem of the pseudoknot are located within the gene for IF3, whereas those forming the 3′ strand are located 280 nt downstream, immediately upstream of the Shine–Dalgarno sequence of the gene for L35. Here we show that premature termination of IF3 translation at a nonsense codon introduced upstream of the pseudoknot results in a substantial enhancement of L20-mediated repression of L35 expression. Conversely, an increase of IF3 translation decreases repression. These results, in addition to an analysis of the effect of mutations in sequences forming the pseudoknot, indicate that IF3 translation decreases L20-mediated repression of L35 expression. We propose that ribosomes translating IF3 disrupt the pseudoknot and thereby attenuate repression. The result is a novel type of translational coupling, where unfolding of the pseudoknot by ribosomes translating IF3 does not increase expression of L35 directly, but alleviates its repression by L20.

Graded transcriptional response to different concentrations of a single transactivator

Threshold mechanisms of transcriptional activation are thought to be critical for translating continuous gradients of extracellular signals into discrete all-or-none cellular responses, such as mitogenesis and differentiation. Indeed, unequivocal evidence for a graded transcriptional response in which the concentration of inducer directly correlates with the level of gene expression in individual eukaryotic cells is lacking. By using a novel binary tetracycline regulatable retroviral vector system, we observed a graded rather than a threshold mechanism of transcriptional activation in two different model systems. When polyclonal populations of cells were analyzed at the single cell level, a dose-dependent, stepwise increase in expression of the reporter gene, green fluorescent protein (GFP), was observed by fluorescence-activated cell sorting. These data provide evidence that, in addition to the generally observed all-or-none switch, the basal transcription machinery also can respond proportionally to changes in concentration of extracellular inducers and trancriptional activators.

Two Yeast La Motif-containing Proteins Are RNA-binding Proteins that Associate with Polyribosomes

We have characterized two Saccharomyces cerevisiae proteins, Sro9p and Slf1p, which contain a highly conserved motif found in all known La proteins. Originally described as an autoantigen in patients with rheumatic disease, the La protein binds to newly synthesized RNA polymerase III transcripts. In yeast, the La protein homologue Lhp1p is required for the normal pathway of tRNA maturation and also stabilizes newly synthesized U6 RNA. We show that deletions in both SRO9 and SLF1 are not synthetically lethal with a deletion in LHP1, indicating that the three proteins do not function in a single essential process. Indirect immunofluorescence microscopy reveals that although Lhp1p is primarily localized to the nucleus, Sro9p is cytoplasmic. We demonstrate that Sro9p and Slf1p are RNA-binding proteins that associate preferentially with translating ribosomes. Consistent with a role in translation, strains lacking either Sro9p or Slf1p are less sensitive than wild-type strains to certain protein synthesis inhibitors. Thus, Sro9p and Slf1p define a new and possibly evolutionarily conserved class of La motif-containing proteins that may function in the cytoplasm to modulate mRNA translation.

Signal recognition particle components in the nucleolus

The signal recognition particle (SRP) is a ribonucleoprotein composed of an Alu domain and an S domain. The S domain contains unique sequence SRP RNA and four SRP proteins: SRP19, SRP54, SRP68, and SRP72. SRP interacts with ribosomes to bring translating membrane and secreted proteins to the endoplasmic reticulum (ER) for proper processing. Additionally, SRP RNA is a member of a family of small nonribosomal RNAs found recently in the nucleolus, suggesting that the nucleolus is more plurifunctional than previously realized. It was therefore of interest to determine whether other SRP components localize to this intranuclear site. In transfected rat fibroblasts, green fluorescent protein fusions of SRP19, SRP68, and SRP72 localized to the nucleolus, as well as to the cytoplasm, as expected. SRP68 also accumulated in the ER, consistent with its affinity for the ER-bound SRP receptor. SRP54 was detected in the cytoplasm as a green fluorescent protein fusion and in immunofluorescence studies, but was not detected in the nucleolus. In situ hybridization experiments also revealed endogenous SRP RNA in the nucleolus. These results demonstrate that SRP RNA and three SRP proteins visit the nucleolus, suggesting that partial SRP assembly, or another unidentified activity of the SRP components, occurs at the nucleolus. SRP54 apparently interacts with nascent SRP beyond the nucleolus, consistent with in vitro reconstitution experiments showing that SRP19 must bind to SRP RNA before SRP54 binds. Our findings support the notion that the nucleolus is the site of assembly and/or interaction between the family of ribonucleoproteins involved in protein synthesis, in addition to ribosomes themselves.

Translation inhibitors stabilize Escherichia coli mRNAs independently of ribosome protection

Translation inhibitors such as chloramphenicol in prokaryotes or cycloheximide in eukaryotes stabilize many or most cellular mRNAs. In Escherichia coli, this stabilization is ascribed generally to the shielding of mRNAs by stalled ribosomes. To evaluate this interpretation, we examine here how inhibitors affect the stabilities of two untranslated RNAs, i.e., an engineered lacZ mRNA lacking a ribosome binding site, and a small regulatory RNA, RNAI. Whether they block elongation or initiation, all translation inhibitors tested stabilized these RNAs, indicating that stabilization does not necessarily reflect changes in packing or activity of translating ribosomes. Moreover, both the initial RNase E-dependent cleavage of RNAI and lacZ mRNA and the subsequent attack of RNAI by polynucleotide phosphorylase and poly(A)-polymerase were slowed. Among various possible mechanisms for this stabilization, we discuss in particular a passive model. When translation is blocked, rRNA synthesis is known to increase severalfold and rRNA becomes unstable. Meanwhile, the pools of RNase E and polynucleotide phosphorylase, which, in growing cells, are limited because these RNases autoregulate their own synthesis, cannot expand. The processing/degradation of newly synthesized rRNA would then titrate these RNases, causing bulk mRNA stabilization.

Importance of anchor genomes for any plant genome project

Progress in agricultural and environmental technologies is hampered by a slower rate of gene discovery in plants than animals. The vast pool of genes in plants, however, will be an important resource for insertion of genes, via biotechnological procedures, into an array of plants, generating unique germ plasms not achievable by conventional breeding. It just became clear that genomes of grasses have evolved in a manner analogous to Lego blocks. Large chromosome segments have been reshuffled and stuffer pieces added between genes. Although some genomes have become very large, the genome with the fewest stuffer pieces, the rice genome, is the Rosetta Stone of all the bigger grass genomes. This means that sequencing the rice genome as anchor genome of the grasses will provide instantaneous access to the same genes in the same relative physical position in other grasses (e.g., corn and wheat), without the need to sequence each of these genomes independently. (i) The sequencing of the entire genome of rice as anchor genome for the grasses will accelerate plant gene discovery in many important crops (e.g., corn, wheat, and rice) by several orders of magnitudes and reduce research and development costs for government and industry at a faster pace. (ii) Costs for sequencing entire genomes have come down significantly. Because of its size, rice is only 12% of the human or the corn genome, and technology improvements by the human genome project are completely transferable, translating in another 50% reduction of the costs. (iii) The physical mapping of the rice genome by a group of Japanese researchers provides a jump start for sequencing the genome and forming an international consortium. Otherwise, other countries would do it alone and own proprietary positions.

Escherichia coli trigger factor is a prolyl isomerase that associates with nascent polypeptide chains.

Correct folding of newly synthesized proteins is proposed to be assisted by molecular chaperones and folding catalysts. To identify cellular factors involved in the initial stages of this process we searched for proteins associated with nascent polypeptide chains. In an Escherichia coli transcription/translation system synthesizing beta-galactosidase we identified a 58-kDa protein which associated with translating ribosomes but dissociated from these ribosomes upon release of nascent beta-galactosidase. N-terminal sequencing identified it as trigger factor, previously implicated in protein secretion. Direct evidence for association of trigger factor with nascent polypeptide chains was obtained by crosslinking. In a wheat germ translation system complemented with E. coli lysates, epsilon-4-(3-trifluoromethyldiazirino)benzoic acid-lysine residues were incorporated into nascent secretory preprolactin and a nonsecretory preprolactin mutant. Trigger factor crosslinked to both types of nascent chains, provided they were ribosome bound. Trigger factor contains key residues of the substrate-binding pocket of FK506-binding protein-type peptidyl-prolyl-cis/trans-isomerases and has prolyl isomerase activity in vitro. We propose that trigger factor is a folding catalyst acting cotranslationally.

Research in speech communication.

Advances in digital speech processing are now supporting application and deployment of a variety of speech technologies for human/machine communication. In fact, new businesses are rapidly forming about these technologies. But these capabilities are of little use unless society can afford them. Happily, explosive advances in microelectronics over the past two decades have assured affordable access to this sophistication as well as to the underlying computing technology. The research challenges in speech processing remain in the traditionally identified areas of recognition, synthesis, and coding. These three areas have typically been addressed individually, often with significant isolation among the efforts. But they are all facets of the same fundamental issue--how to represent and quantify the information in the speech signal. This implies deeper understanding of the physics of speech production, the constraints that the conventions of language impose, and the mechanism for information processing in the auditory system. In ongoing research, therefore, we seek more accurate models of speech generation, better computational formulations of language, and realistic perceptual guides for speech processing--along with ways to coalesce the fundamental issues of recognition, synthesis, and coding. Successful solution will yield the long-sought dictation machine, high-quality synthesis from text, and the ultimate in low bit-rate transmission of speech. It will also open the door to language-translating telephony, where the synthetic foreign translation can be in the voice of the originating talker.

Peptide inhibitors of peptidyltransferase alter the conformation of domains IV and V of large subunit rRNA: a model for nascent peptide control of translation.

Peptides of 5 and 8 residues encoded by the leaders of attenuation regulated chloramphenicol-resistance genes inhibit the peptidyltransferase of microorganisms from the three kingdoms. Therefore, the ribosomal target for the peptides is likely to be a conserved structure and/or sequence. The inhibitor peptides "footprint" to nucleotides of domain V in large subunit rRNA when peptide-ribosome complexes are probed with dimethyl sulfate. Accordingly, rRNA was examined as a candidate for the site of peptide binding. Inhibitor peptides MVKTD and MSTSKNAD were mixed with rRNA phenol-extracted from Escherichia coli ribosomes. The conformation of the RNA was then probed by limited digestion with nucleases that cleave at single-stranded (T1 endonuclease) and double-stranded (V1 endonuclease) sites. Both peptides selectively altered the susceptibility of domains IV and V of 23S rRNA to digestion by T1 endonuclease. Peptide effects on cleavage by V1 nuclease were observed only in domain V. The T1 nuclease susceptibility of domain V of in vitro-transcribed 23S rRNA was also altered by the peptides, demonstrating that peptide binding to the rRNA is independent of ribosomal protein. We propose the peptides MVKTD and MSTSKNAD perturb peptidyltransferase center catalytic activities by altering the conformation of domains IV and V of 23S rRNA. These findings provide a general mechanism through which nascent peptides may cis-regulate the catalytic activities of translating ribosomes.

Discursos e produção de sentidos: experiência da transferência da política do tratamento diretamente observado de curta duração para o controle da tuberculose em Moçambique - África

A estratégia do Tratamento Diretamente Observado de Curta Duração para o controle da tuberculose (DOTS) tem sido usada por vários países do mundo especificamente pelos 22 mais afetados pela doença, que contribuem com cerca de 80% de casos, porém, apesar da estratégia demonstrar a redução de casos e aumentar a adesão dos pacientes ao tratamento, e ter sido implementada em Moçambique desde a década de 1980, a doença continua a ser um problema grave de saúde pública no país. Este estudo tem como objetivo central: analisar a transferência da política do DOTS a partir da visão dos gestores centrais, provinciais e distritais, e profissionais de saúde da província de Nampula. Trata-se de um estudo qualitativo que usa a Análise de Discurso de matriz francesa como seu referencial teórico metodológico que busca a compreensão dos sentidos a partir das condições da produção. Participaram deste estudo 15 profissionais de saúde que ocupavam as posições de gestores, médicos e profissionais de enfermagem e/ou técnicos, e atuavam no Programa nacional de controle de Tuberculose em Moçambique com o mínimo de um ano de experiência, nos meses de maio a agosto de 2014, nos níveis central, provincial (Nampula), e distrital (em oito distritos da província de Nampula). Para a coleta de dados usou-se um roteiro de entrevista semi-estruturado que permitiu explorar os sentidos produzidos. O corpus em análise foi constituído a partir das entrevistas transcritas. Para o auxílio da organização dos dados usou-se o sofware Atlis.ti versão 6. Este estudo teve a aprovação da Comissão Nacional de Bioética e da autorização do Ministro da Saúde de Moçambique. Para a análise dos dados foram identificados quatro blocos discursivos: experiências adotadas na implementação e manutenção do DOTS; estratégias adotadas na implementação e manutenção de DOTS; potencialidades, fragilidades de DOTS no controle da tuberculose e; discursos não buscados pelo roteiro: possíveis soluções. Os sentidos produzidos enfatizam dizeres inscritos em formações discursivas que desconsideram a subjetividade do enfermo; de fragilidades do sistema de saúde, falta de recursos humanos, falta de transporte, baixos salários, insuficiência de laboratórios, distâncias longas entre a moradia do paciente e a unidade sanitária. Ainda os entrevistados entendem como potencialidade a participação dos agentes comunitários e da família no tratamento da doença. Conclui-se que para o controle da tuberculose usando a estratégia DOTS o compromisso do governo deve ser pragmático traduzindo-se em ações concretas como o aumento do financiamento das ações de controle de tuberculose, incluindo as pesquisas, formação dos recursos humanos e, sobretudo, atuar sobre os determinantes socias da saúde

Análisis traductológico de la obra paremiográfica de Charles Cahier, (francés-español)

This thesis contains a translatological analysis of the Spanish proverbs collected by Charles Cahier in Quelque six mille proverbes et aphorismes usuels empruntés à notre âge et aux siècles derniers. Proverbs and other sententious sayings are part of our day-to-day life, and are more or less intensely used according to cultures and their types of speech. They have always existed in every single civilisation. There is, indeed, no denying that the purpose of proverbs is to convey an old experience. They are quoted by major philosophers and writers of all times. As a result of the interest they have raised, books of proverbs have been published for many centuries in a high number of countries. Proverbs can be found everywhere, both in professional and personal settings, or in a conversation between friends. In France, these sayings are more commonly used in literature than in spoken language, whereas in Spain proverbs can be found at all levels of communication. In this regard, it is interesting to compare the translations of international works to detect a number of misunderstandings regarding the interpretation of paremiological elements. This is why translating proverbs is a genuine, complex issue. This thesis, which is aimed at Spanish and French speakers (including native and foreign speakers), has a double application (translatological and linguistic) and falls within the context of translatological and comparative paremiology...

The Need for Professional Translation and Interpretation in School Districts

The language barrier prevents Latino students from experiencing academic success, and prevents Latino parents from participating in their children's education. Through a review of journal articles, research projects, doctoral dissertations, legislation, and books, this project studies the benefits and dangers of various methods of translating and interpreting in the education system, including issues created by language barriers in schools, common methods of translating and interpreting, and legislation addressing language barriers and education. The project reveals that schools use various methods to translate and interpret, including relying on children, school staff and machine translation, although such methods are often problematic and inaccurate. The project also reveals that professional translation and interpretation are superior to the various non-professional methods.

Chaucer's "makyng" of the Romaunt of the Rose

Despite its being the first testimony of Chaucer’s genius, the interest of modern criticism in the Romaunt has mainly focused on the issue of authorship, whereas the efforts to assess this text as a translation have been limited both in their number and in their scope. This paper discusses Chaucer’s translation of the Roman de la Rose, and provides an evaluation of Fragment A from a modern traductological perspective, while taking account of contemporary theoretical positions. First, this article compares the Romaunt with Chaucer’s later translating practice. Second, taking into account that the immediate audience of the Romaunt would have been cognizant of French, this essay considers the pragmatic function of this translation. Finally, I reconstruct some of Chaucer’s decisions in the translation process, and then I present the translation strategies he adopted in order to create an English metapoem which replicated the spirit of the Roman, thus proving the adequacy of English for poetic expression.

COMENEGO (Corpus Multilingüe de Economía y Negocios): a metadiscursive analysis approach

This paper describes a stage in the COMENEGO project, which is creating comparable corpora of Business texts in order to distribute them among translation practitioners so that they can use this resource when translating economic, business or financial texts. This stage consists of discursive analysis of a pilot specialised corpus initially compiled in French and Spanish. Its textual resources are classified in different categories which need to be confirmed so that they can be useful when including them into the virtual platform which will allow users exploit the corpus and filter their searches according to their specific needs. The aim of this paper is to propose a discursive analysis approach based on the concept of ‘metadiscourse’ (Hyland, 2005).

Traducir en tiempos de crisis: estudio comparativo de la traducción francés-español de colocaciones en el lenguaje de los negocios

El presente artículo estudia la traducción de las colocaciones formadas a partir del término crise, aparecidas en un corpus especializado. Tras reseñar algunos trabajos previos sobre lenguaje económico y metáfora en tiempos de crisis, se clasifican las colocaciones originales identificadas en el corpus según diversos conceptos metafóricos. Por último, se analizan las estrategias de traducción y se valoran con el apoyo de un corpus comparable ad hoc. El análisis revela, por una parte, que las metáforas identificadas pueden asociarse a conceptos como, entre otros, alimentos, catástrofes, enfermedades, objetos o pozos, y, por otra parte, que existe una clara tendencia a la traducción literal, especialmente en el caso de las expresiones asociadas a las enfermedades, si bien también se dan en menor medida otras estrategias de traducción. Los resultados del estudio son de utilidad para la enseñanza de la traducción y el lenguaje económico o en la elaboración de repertorios fraseológicos.