CHROMOSOME ARCHITECTURE: COHESION OF TRANSCRIPTIONALLY SILENCED DOMAINS

染色体结构：转录沉默域的凝聚力

• 批准号: 7602100
• 负责人: Marc R. Gartenberg
• 金额: $ 0.56万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7602100
• 关键词: Affinity Chromatography; Architecture; Attention; Centrifugation; Characteristics; Chromatin; Chromosomes; Cis-Acting Sequence; Collaborations; Computer Retrieval of Information on Scientific Projects Database; DNA; DNA Packaging; DNA biosynthesis; Development; Engineering; Funding; Genetic Recombination; Grant; Heterochromatin; Institution; Mass Spectrum Analysis; Mitotic; Nuclear; Proteins; Reagent; Research; Research Personnel; Resources; Role; Saccharomycetales; Site; Source; Structure; United States National Institutes of Health; Work; Yeasts; cohesion; gene repression; in vivo

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Large regions of eukaryotic chromosomes are transcriptionally repressed due to DNA packaging in heterochromatin, a structure that is heritably propagated from one mitotic cycle to the next. Other chromosomal regions are transcriptionally active for brief periods during early development only to be shut-off permanently by repressive structures related to heterochromatin. Work in the Gartenberg lab has focused on mechanisms of heterochromatic gene repression in budding yeast. The hallmark of our approach over the last decade has been the use of site-specific recombination to physically uncouple silenced loci from other chromosomal sequences and activities. This has led to discoveries regarding the role of cis-acting sequences, DNA replication and nuclear localization in silencing. It did not escape our attention that liberating pieces of heterochromatin might facilitate their purification. Previously we developed a differential centrifugation approach to biochemically enrich DNA circles formed by site-specific recombination in vivo. We showed that the isolated material retained characteristics of silenced chromatin. We have since engineered a tandem affinity purification reagent to further purify the rings. In this collaboration with the John Yates lab we aim to identify the protein constituents of yeast heterochromatin by mass spectrometry.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 由于异染色质中的DNA包装，这种真核染色体的大区域被转录抑制，这种结构从一个有丝分裂循环传播到了下一个结构。在早期发育期间，其他染色体区域在短时间内具有转录活性，仅通过与异染色质有关的抑制结构永久关闭。 Gartenberg实验室的工作重点是发芽酵母中异晶基因抑制的机制。在过去十年中，我们方法的标志是使用特定地点重组来从其他染色体序列和活动中物理脱离沉默的基因座。这导致了关于顺式作用序列，DNA复制和核定位置在沉默中的作用的发现。 它并没有避免我们的注意力，即解放异染色质可能有助于其纯化。以前，我们开发了一种差异离心方法，以在体内由位点特异性重组形成的生化富集DNA圆圈。我们表明，孤立的材料保留了沉默的染色质的特征。此后，我们已经设计了一项串联亲和力净化试剂，以进一步净化环。在与John Yates实验室的合作中，我们旨在通过质谱法确定酵母异染色质的蛋白质成分。