SATB1-protein Interaction/function at the BcL2 mbr/MAR

BcL2 mbr/MAR 处的 SATB1 蛋白相互作用/功能

基本信息

批准号：
7279859
负责人：
LINDA K DURRIN
金额：
$ 15.91万
依托单位：
BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-09-01 至 2009-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7279859
关键词：
AT Rich Sequence Appendix Award BCL-2 Protein BCL2 gene Binding Binding Proteins Biochemical Biological Models Cancer Etiology Cell Line Cells Cellular biology Chromatin Chromatin Loop Chromatin Remodeling Factor Chromosomal translocation Cities Complex DNA DNA Repair DNA Sequence DNA-Binding Proteins DNase-I Footprinting Development Disease Elements Enhancers Exons Follicular Lymphoma Foundations Future Genetic Transcription Goals Human IGH@ gene cluster Imaging technology Indium Knowledge acquisition Laboratories Lead Ligation Lymphoma Matrix Attachment Regions Mediating Medical center Mentors Molecular Conformation Mutagenesis Polymerase Chain Reaction Positioning Attribute Protein Binding Proteins RNA Polymerase II RNA Processing Research Scientist Site Staging Structure Techniques Training Transcriptional Regulation Variant Work Yeasts cell type chromatin remodeling in vivo novel therapeutic target thymocyte transcription factor yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): This award will allow the candidate to acquire an additional year of mentored training in techniques of cell biology and imaging technology, and targeted mutagenesis, in the laboratory of Theodore G. Krontiris at the City of Hope National Medical Center. After this period of mentored training, the candidate will remain at the City of Hope in a Research Scientist position, with responsibilities directed primarily towards research. This training and acquisition of knowledge is essential for her future goals directed towards understanding the mechanism of the Bcl2 t(14;18) translocation which is an instigating factor in the onset of follicular lymphoma. Immediately downstream of the Bcl2 major breakpoint region (mbr) is an AT-rich region that binds special AT-rich sequence binding protein l (SATB1), a 'thymocyte-specific', and MAR associating protein. SATB 1 over-expression down-regulates Bcl2 expression. In vivo DNase I footprinting by ligation-mediated PCR (LM-PCR) revealed constitutive and cell-type specific protein binding at the Bcl2 mbr and also an altered DNA helical configuration dependent upon SATB1. More recently, the yeast two-hybrid system (y2h) was used to identify numerous proteins that interact with SATB 1. The purported function(s) of many of these proteins appears logical for MAR-associating proteins (i.e., chromatin remodeling and transcription). Unexpectedly, SATB 1 demonstrated exclusive interaction with SUMO-1, a protein modifying factor, and with additional SUMO-1 enzymatic molecules. We propose to extend our studies as follows: (1) Characterize, through biochemical, biophysical, and immunological approaches, the interaction of SATB1 with SUMO-1 modifying proteins and with select transcription and chromatin remodeling factors. (2) Use targeted mutagenesis of the Bcl2 mbr/MAR to establish which DNA sequences contribute to MAR function. Use these targeted cell lines to establish the contribution of SATB 1 and MARBc12 to specific protein binding, and to chromatin conformation and DNA topology at the Bcl2 mbr/MAR. We anticipate that successful completion of these studies will elucidate mechanisms that contribute to cell-specific protein and DNA functions that lead to disease state.

描述（由申请人提供）：该奖项将使候选人能够在希望之城国家医疗中心的 Theodore G. Krontiris 实验室获得额外一年的细胞生物学和成像技术以及定向诱变技术方面的指导培训。经过这段时间的指导培训后，候选人将继续留在希望之城担任研究科学家，主要负责研究。这种培训和知识获取对于她未来的目标至关重要，即了解 Bcl2 t(14;18) 易位的机制，而 Bcl2 t(14;18) 易位是滤泡性淋巴瘤发病的诱发因素。紧邻 Bcl2 主要断点区域 (mbr) 的下游是一个富含 AT 的区域，它结合特殊的富含 AT 的序列结合蛋白 l (SATB1)，一种“胸腺细胞特异性”和 MAR 关联蛋白。 SATB 1 过度表达会下调 Bcl2 表达。通过连接介导的 PCR (LM-PCR) 进行的体内 DNase I 足迹揭示了 Bcl2 mbr 上的组成型和细胞类型特异性蛋白结合，以及依赖于 SATB1 的改变的 DNA 螺旋构型。最近，酵母双杂交系统 (y2h) 用于鉴定许多与 SATB 1 相互作用的蛋白质。其中许多蛋白质的声称功能对于 MAR 相关蛋白质（即染色质重塑和转录）来说似乎是合乎逻辑的。出乎意料的是，SATB 1 表现出与 SUMO-1（一种蛋白质修饰因子）以及其他 SUMO-1 酶分子的独特相互作用。我们建议将我们的研究扩展如下：（1）通过生化、生物物理和免疫学方法表征 SATB1 与 SUMO-1 修饰蛋白以及与选定的转录和染色质重塑因子的相互作用。 (2) 使用 Bcl2 mbr/MAR 的定向诱变来确定哪些 DNA 序列有助于 MAR 功能。使用这些靶向细胞系来确定 SATB 1 和 MARBc12 对特定蛋白质结合以及 Bcl2 mbr/MAR 上的染色质构象和 DNA 拓扑的贡献。我们预计这些研究的成功完成将阐明导致疾病状态的细胞特异性蛋白质和 DNA 功能的机制。