Characterization of LRRTM and NGR gene families: Expression and functions

Some leucine-rich repeat (LRR) -containing membrane proteins are known regulators of neuronal growth and synapse formation. In this work I characterize two gene families encoding neuronal LRR membrane proteins, namely the LRRTM (leucine-rich repeat, transmembrane neuronal) and NGR (Nogo-66 receptor) families. I studied LRRTM and NGR family member's mRNA tissue distribution by RT-PCR and by in situ hybridization. Subcellular localization of LRRTM1 protein was studied in neurons and in non-neuronal cells. I discovered that LRRTM and NGR family mRNAs are predominantly expressed in the nervous system, and that each gene possesses a specific expression pattern. I also established that LRRTM and NGR family mRNAs are expressed by neurons, and not by glial cells. Within neurons, LRRTM1 protein is not transported to the plasma membrane; rather it localizes to endoplasmic reticulum. Nogo-A (RTN4), MAG, and OMgp are myelin-associated proteins that bind to NgR1 to limit axonal regeneration after central nervous system injury. To better understand the functions of NgR2 and NgR3, and to explore the possible redundancy in the signaling of myelin inhibitors of neurite growth, I mapped the interactions between NgR family and the known and candidate NgR1 ligands. I identified high-affinity interactions between RTN2-66, RTN3-66 and NgR1. I also demonstrate that Rtn3 mRNA is expressed in the same glial cell population of mouse spinal cord white matter as Nogo-A mRNA, and thus it could have a role in myelin inhibition of axonal growth. To understand how NgR1 interacts with multiple structurally divergent ligands, I aimed first to map in more detail the nature of Nogo-A:NgR1 interactions, and then to systematically map the binding sites of multiple myelin ligands in NgR1 by using a library of NgR1 expression constructs encoding proteins with one or multiple surface residues mutated to alanine. My analysis of the Nogo-A:NgR1 -interactions revealed a novel interaction site between the proteins, suggesting a trivalent Nogo-A:NgR1-interaction. Our analysis also defined a central binding region on the concave side of NgR1's LRR domain that is required for the binding of all known ligands, and a surrounding region critical for binding MAG and OMgp. To better understand the biological role of LRRTMs, I generated Lrrtm1 and Lrrtm3 knock out mice. I show here that reporter genes expressed from the targeted loci can be used for maping the neuronal connections of Lrrtm1 and Lrrtm3 expressing neurons in finer detail. With regard to LRRTM1's role in humans, we found a strong association between a 70 kb-spanning haplotype in the proposed promoter region of LRRTM1 gene and two possibly related phenotypes: left-handedness and schizophrenia. Interestingly, the responsible haplotype was linked to phenotypic variability only when paternally inherited. In summary, I identified two families of neuronal receptor-like proteins, and mapped their expression and certain protein-protein interactions. The identification of a central binding region in NgR1 shared by multiple ligands may facilitate the design and development of small molecule therapeutics blocking binding of all NgR1 ligands. Additionally, the genetic association data suggests that allelic variation upstream of LRRTM1 may play a role in the development of left-right brain asymmetry in humans. Lrrtm1 and Lrrtm3 knock out mice developed as a part of this study will likely be useful for schizophrenia and Alzheimer s disease research.

Monimuotoisuus ja esteettömyys : Näkövammaisten asiantuntijoiden työelämäkokemuksia

The purpose of this study is to analyse education, employment, and work-life experiences of visually impaired persons in expert jobs. The empirical data consists of 30 thematic interviews (24 visually impaired persons, 1 family-member of a visually impaired person, 5 persons working with diversity issues), of supplementary articles, and of statistics on the socio-economic status of the visually impaired. The interviewees experiences of education and employment have been analysed by a qualitative method. The analysis has been deepened by reflecting it against the recent discussion on the concept of diversity. The author s methodological choice as a disability researcher has been to treat the interviewees as co-researchers rather than objects of research. Accessibility in its different forms is a prerequisite of diversity in the workplace, and this study examines what kind of accessibility is required by visually impaired professionals. Access to working life depends on the attitudes prejudices and expectations that society has towards a minority group. Social accessibility is connected with internal relationships in the workplace, and achieving social accessibility is a bilateral process. Information technology has revolutionised the visually impaired people s possibilities of accessing information and performing expert tasks. Accessible environment, good mobility skills, and transportation services enable visually impaired employees to get to their workplaces and to navigate there with ease. Integration has raised the level of education and widened the selection of career options for the visually impaired. However, even visually impaired people with academic degrees often need employment support services. Visually impaired professionals are mainly employed in the public and third sector. Achieving diversity in the labour market is a multiactor process. Social support services are needed, as well as courage and readiness from employers to hire people with disabilities. The organisations of the visually impaired play an important role in affecting the attitudes and providing peer support. Visually impaired employees need good professional skills, blindness skills, and social courage, and they need to be comfortable with their disability. In the workplace, diversity may actualise as diverse ways of working: the work is done by using technical aids or other means of compensating for the lack of eyesight. When an employee must find compensatory solutions for disability-related limitations at work, this will also develop his/her problem-solving abilities. Key words: visually impaired, diversity, accessibility, working life

Eron aika : Tyttärien kertomuksia ikääntyneen vanhemman kuolemasta

There is a relative absence of sociological and cultural research on how people deal with the death of a family member in the contemporary western societies. Research on this topic has been dominated by the experts of psychology, psychiatry and therapy, who mention the social context only in passing, if at all. This gives an impression that the white westerners bereavement experience is a purely psychological phenomenon, an inner journey, which follows a natural, universal path. Yet, as Tony Walter (1999) states, ignoring the influence of culture not only impoverishes the understanding of those work with bereaved people, but it also impoverishes sociology and cultural studies by excluding from their domain a key social phenomenon. This study explores the cultural dimension of grief through narratives told by fifteen of recently bereaved Finnish women. Focussing on one sex only, the study rests on the assumption of the gendered nature of bereavement experience. However, the aim of the study is not to pinpoint the gender differences in grief and mourning, but to shed light on women s ways of dealing with the loss of a loved one in a social context. Furthermore, the study focuses on a certain kind of loss: the death of an elderly parent. Due to the growth in the life expectancy rate, this has presumably become the most typical type of bereavement in contemporary, ageing societies. Most of population will face the death of a parent as they reach the middle years of the life course. The data of this study is gathered with interviews, in which the interviewees were invited to tell a narrative of their bereavement. Narrative constitutes a central concept in this study. It refers to a particular form of talk, which is organised around consequential events. But there are also other, deeper layers that have been added to this concept. Several scholars see narratives as the most important way in which we make sense of experience. Personal narratives provide rich material for mapping the interconnections between individual and culture. As a form of thought, narrative marries singular circumstances with shared expectations and understandings that are learned through participation in a specific culture (Garro & Mattingly 2000). This study attempts to capture the cultural dimension of narrative with the concept of script , which originates in cognitive science (Schank & Abelson 1977) and has recently been adopted to narratology (Herman 2002). Script refers to a data structure that informs how events usually unfold in certain situations. Scripts are used in interpreting events and representing them verbally to others. They are based on dominant forms of knowledge that vary according to time and place. The questions that were posed in this study are the following. What kind of experiences bereaved daughters narrate? What kind of cultural scripts they employ as they attempt to make sense of these experiences? How these scripts are used in their narratives? It became apparent that for the most of the daughters interviewed in this study the single most important part of the bereavement narrative was to form an account of how and why the parent died. They produced lengthy and detailed descriptions of the last stage of a parent s life in contrast with the rest of the interview. These stories took their start from a turn in the parent s physical condition, from which the dying process could in retrospect be seen to have started, and which often took place several years before the death. In addition, daughters also talked about their grief reactions and how they have adjusted to a life without the deceased parent. The ways in which the last stage of life was told reflect not only the characteristic features of late modernity but also processes of marginalisation and exclusion. Revivalist script and medical script, identified by Clive Seale as the dominant, competing models for dying well in the late modern societies, were not widely utilised in the narratives. They could only be applied in situations in which the parent had died from cancer and at somewhat younger age than the average. Death that took place in deep old age was told in a different way. The lack of positive models for narrating this kind of death was acknowledged in the study. This can be seen as a symptom of the societal devaluing of the deaths of older people and it affects also daughters accounts of their grief. Several daughters told about situations in which their loss, although subjectively experienced, was nonetheless denied by other people.

mRNA-binding protein HuR in breast carcinogenesis

Breast cancer is the most common cancer among women. Although its prognosis has improved nowadays, methods to predict the progression of the disease or to treat it are not comprehensive. This thesis work was initiated to elucidate in breast carcinogenesis the role of HuR, a ubiquitously expressed mRNA-binding protein that regulates gene expression posttranscriptionally. HuR is predominantly nuclear, but it shuttles between the nucleus and the cytoplasm, and this nucleocytoplasmic translocation is important for its function as a RNA-stabilizing and translational regulator. HuR has been associated with diverse cellular processes, for example carcinogenesis. The specific aims of my thesis work were to study the prognostic value of HuR in breast cancer and to clarify the mechanisms by which HuR contributes to breast carcinogenesis. My ultimate goal is, by better understanding the role of HuR in breast carcinogenesis, to aid in the discovery of novel targets for cancer therapies. HuR expression and localization was studied in paraffin-embedded preinvasive (atypical ductal hyperplasia, ADH, and ductal carcinoma in situ, DCIS) specimens as well in sporadic and familial breast cancer specimens. Our results show that cytoplasmic HuR expression was already elevated in ADH and remained elevated in DCIS as well as in cancer specimens. Clinicopathological analysis showed that cytoplasmic HuR expression associated with the more aggressive form of the disease in DCIS, and in cancer specimens it proved an independent marker for poor prognosis. Importantly, cytoplasmic HuR expression was significantly associated with poor outcome in the subgroups of small (2 cm) and axillary lymph node-negative breast cancers. HuR proved to be the first mRNA stability protein the expression of which is associated in breast cancer with poor outcome. To explore the mechanisms of HuR in breast carcinogenesis, lentiviral constructs were developed to inhibit and to overexpress the HuR expression in a breast epithelial cell line (184B5Me). Our results suggest that HuR mediates breast carcinogenesis by participating in processes important in cell transformation, in programmed cell death, and in cell invasion. Global gene expression analysis shows that HuR regulates genes participating in diverse cellular processes, and affects several pathways important in cancer development. In addition, we identified two novel target transcripts (connective tissue growth factor, CTGF, and Ras oncogene family member 31, RAB31) for HuR. In conclusion, because cytoplasmic HuR expression in breast cancer can predict the outcome of the disease it could serve in clinics as a prognostic marker. HuR accumulates in the cytoplasm even at its non-invasive stage (ADH and DCIS) of the carcinogenic process and supports functions essential in cell alteration. These data suggest that HuR contributes to carcinogenesis of the breast epithelium.

Effects of gender, and SLCO1B1 and CYP7A1 polymorphisms on plasma bile acids in humans and the pharmacokinetics of therapeutic ursodeoxycholic acid

Bile acids are important steroid-derived molecules essential for fat absorption in the small intestine. They are produced in the liver and secreted into the bile. Bile acids are transported by bile flow to the small intestine, where they aid the digestion of lipids. Most bile acids are reabsorbed in the small intestine and return to the liver through the portal vein. The whole recycling process is referred to as the enterohepatic circulation, during which only a small amount of bile acids are removed from the body via faeces. The enterohepatic circulation of bile acids involves the delicate coordination of a number of bile acid transporters expressed in the liver and the small intestine. Organic anion transporting polypeptide 1B1 (OATP1B1), encoded by the solute carrier organic anion transporter family, member 1B1 (SLCO1B1) gene, mediates the sodium independent hepatocellular uptake of bile acids. Two common SNPs in the SLCO1B1 gene are well known to affect the transport activity of OATP1B1. Moreover, bile acid synthesis is an important elimination route for cholesterol. Cholesterol 7α-hydroxylase (CYP7A1) is the rate-limiting enzyme of bile acid production. The aim of this thesis was to investigate the effects of SLCO1B1 polymorphism on the fasting plasma levels of individual endogenous bile acids and a bile acid synthesis marker, and the pharmacokinetics of exogenously administered ursodeoxycholic acid (UDCA). Furthermore, the effects of CYP7A1 genetic polymorphism and gender on the fasting plasma concentrations of individual endogenous bile acids and the bile acid synthesis marker were evaluated. Firstly, a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the determination of bile acids was developed (Study I). A retrospective study examined the effects of SLCO1B1 genetic polymorphism on the fasting plasma concentrations of individual bile acids and a bile acid synthesis marker in 65 healthy subjects (Study II). In another retrospective study with 143 healthy individuals, the effects of CYP7A1 genetic polymorphism and gender as well as SLCO1B1 polymorphism on the fasting plasma levels of individual bile acids and the bile acid synthesis marker were investigated (Study III). The effects of SLCO1B1 polymorphism on the pharmacokinetics of exogenously administered UDCA were evaluated in a prospective genotype panel study including 27 healthy volunteers (Study IV). A robust, sensitive and simple HPLC-MS/MS method was developed for the simultaneous determination of 16 individual bile acids in human plasma. The method validation parameters for all the analytes met the requirements of the FDA (Food and Drug Administration) bioanalytical guidelines. This HPLC-MS/MS method was applied in Studies II-IV. In Study II, the fasting plasma concentrations of several bile acids and the bile acid synthesis marker seemed to be affected by SLCO1B1 genetic polymorphism, but these findings were not replicated in Study III with a larger sample size. Moreover, SLCO1B1 polymorphism had no effect on the pharmacokinetic parameters of exogenously administered UDCA. Furthermore, no consistent association was observed between CYP7A1 genetic polymorphism and the fasting plasma concentrations of individual bile acids or the bile acid synthesis marker. In contrast, gender had a major effect on the fasting plasma concentrations of several bile acids and also total bile acids. In conclusion, gender, but not SLCO1B1 or CYP7A1 polymorphisms, has a major effect on the fasting plasma concentrations of individual bile acids. Moreover, the common genetic polymorphism of CYP7A1 is unlikely to influence the activity of CYP7A1 under normal physiological conditions. OATP1B1 does not play an important role in the in vivo disposition of exogenously administered UDCA.

Interactions of natural products with β-lactoglobulins, members of the lipocalin family

Natural products constitute an important source of new drugs. The bioavailability of the drugs depends on their absorption, distribution, metabolism and elimination. To achieve good bioavailability, the drug must be soluble in water, stable in the gastrointestinal tract and palatable. Binding proteins may improve the solubility of drug compounds, masking unwanted properties, such as bad taste, bitterness or toxicity, transporting or protecting these compounds during processing and storage. The focus of this thesis was to study the interactions, including ligand binding and the effect of pH and temperature, of bovine and reindeer β-lactoglobulin (βLG) with such compounds as retinoids, phenolic compounds as well as with compounds from plant extracts, and to investigate the transport properties of the βLG-ligand complex. To examine the binding interactions of different ligands to βLG, new methods were developed. The fluorescence binding method for the evaluation of ligand binding to βLG was miniaturized from a quartz cell to a 96-well plate. A method of ultrafiltration sampling combined with high-performance liquid chromatography was developed to assess the binding of compounds from extracts. The interactions of phenolic compounds or retinoids and βLG were investigated using the 96-well plate method. The majority of flavones, flavonols, flavanones and isoflavones and all of the retinoids included were shown to bind to bovine and reindeer βLG. Phenolic compounds, contrary to retinol, were not released at acidic pH. Those results suggest that βLG may have more binding sites, probably also on the surface of βLG. An extract from Camellia sinensis (L.) O. Kunze (black tea), Urtica dioica L. (nettle) and Piper nigrum (black pepper) were used to evaluate whether βLG could bind compounds from plant extracts. Piperine from P. nigrum was found to bind tightly and rutin from U. dioica weakly to βLG. No components from C. sinensis bound to βLG in our experiment. The uptake and membrane permeation of bovine and reindeer βLG, free and bound with retinol, palmitic acid and cholesterol, were investigated using Caco-2 cell monolayers. Both bovine and reindeer βLG were able to cross the Caco-2 cell membrane. Free and βLG-bound retinol and palmitic acid were transported equally, whereas cholesterol could not cross the Caco-2 cell monolayer free or bound to βLG. Our results showed that βLG can bind different natural product compounds, but cannot enhance transport of retinol, palmitic acid or cholesterol through Caco-2 cells. Despite this, βLG, as a water-soluble binding protein, may improve the solubility of natural compounds, possibly protecting them from early degradation and transporting some of them through the stomach. Furthermore, it may decrease their bad or bitter taste during oral administration of drugs or in food preparations. βLG can also enhance or decrease the health benefits of herbal teas and food preparations by binding compounds from extracts.

Murtuva säätyvalta, kestävä eliitti : Senaattori Lennart Gripenbergin sukupiiri ja sääty-yhteiskunnan muodonmuutos

This study examines the transformation of the society of estates in the Finnish Grand Duchy through the case study of Senator Lennart Gripenberg and his family circle. While national borders and state structures changed, the connections between old ruling elite families remained intact as invisible family networks, ownership relations, economic collaboration and power of military families. These were the cornerstones of trust, which helped to strengthen positions gained in society. Also, these connections often had a central if unperceivable impact on social development and modernization. Broadly speaking, the intergenerational social reproduction made it possible for this network of connections to remain in power and, as an imperceptible factor, also influenced short-term developments in the long run. Decisions which in the short term appeared unproductive, would in the long run produce cumulative immaterial and material capital across generations as long-term investments. Social mobility, then, is a process which clearly takes several generations to become manifest. The study explores long-term strategies of reproducing and transferring the capital accumulated in multinational elite networks. Also, what was the relationship of these strategies to social change? For the representatives of the military estate the nobility and for those men of the highest estates who had benefited from military training, this very education of a technical-military nature was the key to steering, controlling and dealing with the challenges following the industrial breakthrough. The disintegration of the society of estates and the rising educational standards also increased the influence of those professionals previously excluded, which served to intensify competition for positions of power. The family connections highlighted in this study overlapped in many ways, working side by side and in tandem to manage the economic and political life in Finland, Russia and Sweden. The analysis of these ties has opened up a new angle to economic co-operation, for example, as seen in the position of such family networks not only in Finnish, but also Swedish and Russian corporations and in the long historical background of the collaboration. This also highlights in a new way the role of women in transferring the cumulative social capital and as silent business partners. The marriage strategies evident in business life clearly had an impact on the economic life. The collaborative networks which transcended generations, national boundaries and structures also uncover, as far as the elites are concerned, serious problems in comparative studies conducted from purely national premises. As the same influential families and persons in effect held several leading positions in society, the line would blur between public and invisible uses of power. The power networks thus aimed to build monopolies to secure their key positions at the helm. This study therefore examines the roles of Lennart Gripenberg senator, business executive, superintendent of the Department of Industry, factory inspector, and founding member of industrial interest groups as part of the reproduction strategies of the elite. The family and other networks of the powerful leaders of society, distinguished by social, economic and cultural capital, provided a solid backdrop for the so-called old elites in their quest for strategies to reproducing power in a changing world. Crucially, it was easier for the elites to gain expertise to steer the modernization process and thereby secure for the next generation a leading position in society, something that they traditionally, too, had had the greatest interest in.

Tohtoriksi tulemisen tarina

The study examines the dissertation process. The thesis is based on twenty-three autobiographical stories. They were collected from PhDs who had taken their doctoral degrees at the University of Helsinki. The purpose is to investigate the experiences of these PhD recipients and find out what they recall of the events, traditions, and meanings associated with their PhD studies. An anthology collected from PhDs in the historical sciences, which was published in 1998, was used as reference material. This study begins with a general presentation of the history and values of the university and the traditions of the academic community in Finland. Thereafter, the data and the methodological framework of the study are presented. Attention is paid to the discipline background, sex and age of these PhDs, which may have affected their storytelling. The former studies concerning the academic life together with the experiences of the writers revealed that the process of becoming a PhD graduate is generally considered as an academic rite of passage. The change from student status to becoming a fully accepted professional member of one’s field is real, transforming and at certain points, ritualized. The finding of an inner logic to the doctoral process, which included meaningful turning points, was central to the narrative analysis approach. During the process there were different kinds of struggles and highlights. The family background, university studies and time in employment had directed the doctoral studies of some PhD students enormously. But generally the most memorable and value laden were the phases of actually writing the dissertation, the public examination, and the celebration party in the evening called “karonkka”. The picture that emerged from these various life-stories demonstrated that there was no ivory tower where doctoral candidates live an isolated existence. Everyday cares in ordinary life and tasks in academic life are both demanding and rewarding at the same time. The main point is that the academic community in general and PhD supervisors in particular should concentrate not only on the thesis-writing process their students but also on the doctoral students´ wider life situation. Doctoral students need scientific, financial, and mental support. Somebody must be available to inspire and offer encouragement otherwise the process will be disturbed. The successful completion of the PhD ritual gives the PhD graduate self-confidence and respect but its’ influence on the doctors’ subsequent academic career is diverse. The dissertation process is a personal and unique experience. The aim of the PhD graduation is that this traditional ritual leads to a successful completion and ensures a positive reward for the PhD graduate and the academic community. Keywords: autobiography, narratives, academic traditions, PhD graduates.

Intergenerational Transfer of Sabbath Observance : How is Religious Tradition Transferred to Children among Finnish Seventh-day Adventists?

This study examined religious home education in educational, psychological, and sociological context. Growing up within a religious denomination is a process of learning the rules, norms, opinions, and attitudes, which serve to make the individual an active member of the group. It is a process of transferring the cultural inheritance between generations. Sabbathkeeping can be regarded as a strong indicator of the Seventh-day Adventist value system, which is also why I have concentrated on this specific issue in my study. The purpose of the study was to find out, how the Sabbath is transferred from parents to children among Finnish Adventists. It was also examined how parents could make the day of rest positively exceptional for children, and how the parental authoritativeness affects the process of transference. According to Bull & Lockhart s (1989) theory, the amount of Adventist generations in family history influences the transfer of religious tradition. This study aimed to find out whether or not this theory would apply to the present-day Finland. The nature of religious development among Adventist young people was also one of the interests of the research. The methods used in the study were in-depth interviews (n = 10) and a survey (n = 106). The majority of the interviewees was young adults (age 15-30) grown up in Adventist families. The interviews were taped and transcribed for the study, and survey answers were analysed with SPSS-data analysis program. The amount of survey questionnaires evaluated was 106, whole population of 15-30 year-old Finnish Adventists being about one thousand. Democratic relationship between parents and children, parents' example, encouragement to own thinking, and positive experiences of Sabbath and the whole religion, including the social dimension of the Adventism, seem to be some of the most significant factors in transference of religious tradition. Both too severe and too permissive education were considered to lead to similar results: unsuccessful transfer of values, or even rebellion and adopting a totally opposite way of life than that of the parents. In this study the amount of Adventist generations in family history does not correlate significantly with the end results of value transference. Keywords: Sabbath, intergenerational, value transference, religious home education Avainsanat: sapatti, arvojen siirtyminen vanhemmilta lapsille, uskonnollinen kotikasvatus

Molecular Mechanisms of Androgen Receptor Interactions

The androgen receptor (AR) mediates the effects of the male sex-steroid hormones (androgens), testosterone and 5?-dihydrotestosterone. Androgens are critical in the development and maintenance of male sexual characteristics. AR is a member of the steroid receptor ligand-inducible transcription factor family. The steroid receptor family is a subgroup of the nuclear receptor superfamily that also includes receptors for the active forms of vitamin A, vitamin D3, and thyroid hormones. Like all nuclear receptors, AR has a conserved modular structure consisting of a non-conserved amino-terminal domain (NTD), containing the intrinsic activation function 1, a highly conserved DNA-binding domain, and a conserved ligand-binding domain (LBD) that harbors the activation function 2. Each of these domains plays an important role in receptor function and signaling, either via intra- and inter-receptor interactions, interactions with specific DNA sequences, termed hormone response elements, or via functional interactions with domain-specific proteins, termed coregulators (coactivators and corepressors). Upon binding androgens, AR acquires a new conformational state, translocates to the nucleus, binds to androgen response elements, homodimerizes and recruits sequence-specific coregulatory factors and the basal transcription machinery. This set of events is required to activate gene transcription (expression). Gene transcription is a strictly modulated process that governs cell growth, cell homeostasis, cell function and cell death. Disruptions of AR transcriptional activity caused by receptor mutations and/or altered coregulator interactions are linked to a wide spectrum of androgen insensitivity syndromes, and to the pathogenesis of prostate cancer (CaP). The treatment of CaP usually involves androgen depletion therapy (ADT). ADT achieves significant clinical responses during the early stages of the disease. However, under the selective pressure of androgen withdrawal, androgen-dependent CaP can progress to an androgen-independent CaP. Androgen-independent CaP is invariably a more aggressive and untreatable form of the disease. Advancing our understanding of the molecular mechanisms behind the switch in androgen-dependency would improve our success of treating CaP and other AR related illnesses. This study evaluates how clinically identified AR mutations affect the receptor s transcriptional activity. We reveal that a potential molecular abnormality in androgen insensitivity syndrome and CaP patients is caused by disruptions of the important intra-receptor NTD/LBD interaction. We demonstrate that the same AR LBD mutations can also disrupt the recruitment of the p160 coactivator protein GRIP1. Our investigations reveal that 30% of patients with advanced, untreated local CaP have somatic mutations that may lead to increases in AR activity. We report that somatic mutations that activate AR may lead to early relapse in ADT. Our results demonstrate that the types of ADT a CaP patient receives may cause a clustering of mutations to a particular region of the receptor. Furthermore, the mutations that arise before and during ADT do not always result in a receptor that is more active, indicating that coregulator interactions play a pivotal role in the progression of androgen-independent CaP. To improve CaP therapy, it is necessary to identify critical coregulators of AR. We screened a HeLa cell cDNA library and identified small carboxyl-terminal domain phosphatase 2 (SCP2). SCP2 is a protein phosphatase that directly interacts with the AR NTD and represses AR activity. We demonstrated that reducing the endogenous cellular levels of SCP2 causes more AR to load on to the prostate specific antigen (PSA) gene promoter and enhancer regions. Additionally, under the same conditions, more RNA polymerase II was recruited to the PSA promoter region and overall there was an increase in androgen-dependent transcription of the PSA gene, revealing that SCP2 could play a role in the pathogenesis of CaP.