Impact of chromatin structure on alternative splicing of CD45 pre-mRNA

染色质结构对 CD45 前体 mRNA 选择性剪接的影响

• 批准号: 8175355
• 负责人: Shalini Oberdoerffer
• 金额: $ 37.55万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8175355
• 关键词: Affect; Alternative Splicing; Area; B-Lymphocytes; Binding; Biological Models; Cell Line; Cell model; Chromatin; Chromatin Structure; DNA; DNA Binding; DNA Polymerase II; Data; Exons; Flow Cytometry; Genes; Goals; Mediating; PTPRC gene; Postdoctoral Fellow; Process; Protein Isoforms; RNA Splicing; Relative (related person); Ribonucleoproteins; Structure; System; Technology; Transcript; cell type; genome-wide; mRNA Precursor

As a postdoctoral fellow, I showed that the heterogeneous ribonucleoprotein L-like (hnRNPLL) was a master regulator of the transition from the CD45RA to CD45RO isoform. I further identified hnRNPLL-expressing BJAB B cells to exclusively express the CD45RO isoform, whereas the hnRNPLL-negative BL41 B cell line exclusively expressed the CD45RA isoform at steady state. I am presently using these cellular model systems to identify differences in chromatin structure that correlate with differential CD45 isoform expression. Our preliminary data show that there is very little difference in the composition of the DNA in both cell types. However, we do find areas of relative enrichment of DNA binding factors, suggesting that chromatin acts to poise the DNA in a particular structure that influences pol II-mediated splicing decisions. These studies have utilized technologies such as Chromatin-immunopreciptiation, lentiviral transduction and flow cytometry.

作为博士后研究员，我表明异质核糖核蛋白L样（HNRNPLL）是从CD45RA到CD45RO同工型的过渡的主要调节剂。我进一步鉴定了表达HNRNPLL的BJAB B细胞，以专门表达CD45RO同工型，而HNRNPLL阴性BL41 B细胞系在稳态下仅表达CD45RA同工型。我目前正在使用这些细胞模型系统来识别与差异CD45同工型表达相关的染色质结构的差异。 我们的初步数据表明，在两种细胞类型中，DNA的组成几乎没有差异。但是，我们确实找到了DNA结合因子相对富集的领域，这表明染色质起作用以影响Pol II介导的剪接决策的特定结构中的DNA。 这些研究利用了诸如染色质免疫沉淀，慢病毒转导和流式细胞仪等技术。