Biochemical Studies of a Transcription Factor

转录因子的生化研究

• 批准号: 7665133
• 负责人: MICHAEL Aaron WEISS
• 金额: $ 31.42万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7665133
• 关键词: Abbreviations; Address; Admixture; Adult; Alleles; Animals; Behavior; Behavioral; Behavioral Genetics; Binding; Biochemical; Biochemistry; Biological; Biological Process; C-terminal; Caenorhabditis elegans; Chromosomes, Human, Pair 9; Co-Immunoprecipitations; Collaborations; Complement component C1s; Complex; Congenital Abnormality; Coupled; Courtship; DNA; DNA Binding; DNA Sequencing Facility; Databases; Development; Developmental Biology; Dimerization; Disease; Drosophila genus; Drosophila melanogaster; Elements; Exhibits; Family; Female; Foundations; Gene Expression Regulation; Gene Targeting; Genes; Genetic; Genetic Transcription; Goals; Hand; Homeobox Genes; Human; Human Chromosomes; Hybrids; In Vitro; Indium; Infertility; Laboratories; Location; Mass Spectrum Analysis; Measurement; Mediating; Methods; Modeling; Molecular; Molecular Genetics; Mutation; N-terminal; Nervous system structure; Newborn Infant; Nuclear Magnetic Resonance; Phenotype; Property; Protein Binding; Protein Isoforms; Proteins; Proteomics; Protons; RNA Splicing; Regulation; Research; Resolution; Screening procedure; Sexual Development; Signal Transduction; Site; Specific qualifier value; Staging; Structure; Structure-Activity Relationship; Syndrome; System; Tail; Technology; Temperature; Tissues; Transcription Coactivator; Transcription Factor 3; Transgenes; Transgenic Organisms; Ubiquitin; Validation; Variant; Work; Yeasts; base; design; dimer; fly; gel mobility shift assay; genetic analysis; gonadal cancer; in vivo; innovation; insight; intercellular communication; loss of function; male; member; neural circuit; nuclear Overhauser enhancement; null mutation; programs; protein function; protein protein interaction; protein structure; public health relevance; sex; sex determination; sexual dimorphism; structural biology; trait; transcription factor; yeast two hybrid system

DESCRIPTION (provided by applicant): Development in metazoans is regulated by specific regulatory hierarchies leading to the stage- and tissue-specific expression of transcriptional networks. Understanding such networks poses a major challenge in developmental biology. A model is provided by the regulation of sexual dimorphism in Drosophila melanogaster. This collaborative R01 application from the laboratories of M. Weiss (CWRU) and B. Baker (Stanford) focuses on Doublesex (DSX), a member of a newly recognized family of DM transcription factors broadly involved in metazoan development, including in humans. Our long-term goal is a molecular understanding of how information is transmitted from the primary determinants of sex to the genes responsible for the morphological, biochemical, and behavioral differences between the sexes. The doublesex (dsx) gene controls a major branch of the sex-determining hierarchy of D. melanogaster. The doublesex (dsx) gene encodes sex-specific protein isoforms (designated DSXF (female) and DSXM (male)) as a consequence of sex-specific RNA splicing. Although DSXF and DSXM have opposing biological functions, they exhibit similar DNA-binding properties and so presumably recognize the same cis- acting control elements in target genes. DSXF (but not DSXM) interacts with transcriptional coactivator Intersex (IX), presumably leading to assembly of a female-specific multiprotein-DNA complex at target genes. Such sex-specific regulatory properties presumably underlie the function of dsx as a prototypical "behavioral gene" in the nervous system: DSXM regulates sine singing, a component of the male courtship song, whereas DSXF is hypothesized to regulate major features of female courtship behavior, including rejection behavior. We seek to investigate the molecular bases of the sex-specific functions of DSXF and DSXM with application to behavioral genetics. To these ends, an innovative interdisciplinary strategy is proposed that integrates molecular genetics with biochemistry, proteomics, and structural biology. Studies will focus on the respective C-terminal domains of DSXF/M containing sex-specific tails. In addition, newly described methods for gene targeting in vivo will be employed to investigate the relationship between dsx as a behavioral gene and a discrete and quantifiable repertoire of behavioral elements. Because deletions of DSX-related genes in human chromosome 9 are associated with intersexual abnormalities of the newborn, our results may also provide insight into a major class of human birth defects. Public Health Relevance: How the differences between male and female animals are specified by genes defines a major scientific problem. We are studying sex-specific genes in fruit flies to gain insight into human birth defects, infertility syndromes, and cancers of the gonads.

描述（由申请人提供）：后生动物的发展受到特定的调节层次结构的调节，导致转录网络的阶段和组织特异性表达。了解此类网络在发育生物学方面构成了重大挑战。通过调节果蝇中的性二态性的调节提供了模型。 M. Weiss（CWRU）和B. Baker（Stanford）实验室的合作R01应用集中在Doublesex（DSX）上，Doublesex（DSX）是新认可的DM转录因子家族的成员，广泛地参与了内唑发展，包括人类。我们的长期目标是对信息从性别的主要决定因素传递到负责形态学，生化和性别之间行为差异的基因的分子理解。 Doublesex（DSX）基因控制了D. Melanogaster的性别确定层次结构的主要分支。 Doublesex（DSX）基因编码性别特异性的同工型（指定的DSXF（雌性）和DSXM（雄性）），这是由于性别特异性RNA剪接的结果。尽管DSXF和DSXM具有相对的生物学功能，但它们具有相似的DNA结合特性，因此大概可以识别靶基因中相同的表达控制元件。 DSXF（而非DSXM）与转录共激活因子互联（IX）相互作用，大概会导致靶基因上女性特异性多蛋白DNA复合物的组装。这种性别特异性的调节特性大概是DSX作为神经系统中原型“行为基因”的功能的基础：DSXM调节正弦唱歌，这是男性求爱歌曲的组成部分，而DSXF则被认为可以调节女性求爱行为的主要特征，包括拒绝行为。我们试图研究DSXF和DSXM性别特异性功能的分子碱基，并应用于行为遗传学。为此，提出了一种创新的跨学科策略，该策略将分子遗传学与生物化学，蛋白质组学和结构生物学相结合。研究将集中于含有性别特异性尾巴的DSXF/m的各个C末端结构域。此外，将采用新描述的针对体内基因的方法来研究DSX作为行为基因的关系以及行为元素的离散且可量化的曲目。由于人类染色体中与DSX相关的基因的缺失与新生儿的两性异常有关，因此我们的结果也可能会洞悉主要的人类先天缺陷。 公共卫生相关性：基因指定男性和雌性动物之间的差异如何定义一个主要的科学问题。我们正在研究果蝇中的性别特异性基因，以深入了解人类的先天缺陷，不育综合征和性腺癌。