Analysis of the yeast SWI1, SWI2, and SWI3 proteins

酵母 SWI1、SWI2 和 SWI3 蛋白的分析

• 批准号: 8306813
• 负责人: Craig L Peterson
• 金额: $ 54.44万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8306813
• 关键词: ATP Hydrolysis; Acetylation; Affect; Binding; Biochemical; Biochemistry; Biological; Biological Assay; Cells; Chromatin; Chromosome Structures; Collaborations; Complex; DNA; DNA Sequence; DNA Transposable Elements; DNA lesion; Development; Dinucleosome; Enzymes; Event; Excision; Gene Expression; Genes; Genetic; Genetic Transcription; Heterochromatin; Histone Acetylation; Image; In Vitro; Instruction; Iowa; Journals; Kinetics; Lead; Location; Malignant Neoplasms; Maps; Mediating; Methodology; Molecular Conformation; Molecular Genetics; Mono-S; Mouse Mammary Tumor Virus; Nuclear; Nucleosomes; Outcome; Plasmids; Play; Process; Proteins; Publishing; Reaction; Research; Resolution; Role; SAGA; SMARCA1 gene; SWI1; Site; Slide; Staining method; Stains; Structure; System; Testing; Transcription Coactivator; Universities; Work; Yeasts; analog; base; dimer; fusion gene; gene repression; hSWI/SNF; homologous recombination; in vivo; novel; preference; prevent; protein complex; recombinational repair; research study; single molecule

The overall objective of our research is to determine how chromosome structure affects gene expression and how the transcription machinery contends with this structure. Our general strategy has been to focus on an evolutionarily conserved protein complex, SWI/SNF, which is required for expression of a subset of yeast genes and for the activity of several transcriptional activators. Genetic studies in yeast indicate that SWI/SNF facilitates transcription by antagonizing chromatin-mediated transcriptional repression, and our in vitro studies indicate that the ~1 Mda SWI/SNF complex can use the energy derived from ATP hydrolysis to mobilize nucleosomes and disrupt nucleosome structure. Our recent work indicates that SWI/SNF may be unique among remodeling enzymes as its action can disrupt compact heterochromatin, promoting homologous recombination. This proposal continues to exploit the powerful genetic and biochemical opportunities available in yeast to investigate the role of SWI/SNF in vivo and the biochemical mechanism by which SWI/SNF disrupts euchromatic and heterochromatic structures in vitro. Our first objective will dissect the biochemical mechanism by which SWI/SNF remodels heterochromatin structures, including experiments that test whether SWI/SNF contains a unique subunit that interacts with heterochromatin components. In our second objective we will investigate whether SWI/SNF plays a general role in heterochromatin dynamics in vivo. We propose to test if SWI/SNF disrupts the spurious assembly of heterochromatic structures at euchromatic loci, and whether SWI/SNF facilitates Ty5 transposition into heterochromatin. Our third objective will investigate the role of DNA sequence and histone acetylation in regulating the type of remodeling produced due to SWI/SNF action. This aim involves chromatin biochemistry and single molecule approaches. In our fourth aim, we propose to investigate the structure of SWI/SNF and SWI/SNF- nucleosome complexes by EM methodologies. These studies also involve imaging of complexes that harbor subunit-GFP fusions to map locations of SWI/SNF subunits within the obtained structures.

我们研究的总体目标是确定染色体结构如何影响基因表达和