Characterization of candidate histone methyltransferases

候选组蛋白甲基转移酶的表征

• 批准号: 7338646
• 负责人: David Eric Symer
• 金额: --
• 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/7338646
• 关键词: Characterization; candidate; histone; methyltransferases

We conducted a comprehensive analysis of transcription of a candidate histone methyltransferase, Setdb2, in human and mouse tissues, revealing tissue-specific expression of alternatively spliced and chimeric transcripts. We found in-frame fusion transcripts linking Setdb2 to an adjacent independent gene, Phf11, which encodes a zinc finger-containing plant homeodomain motif. We previously found several protein-binding partners for the putative histone methyltransferases using yeast two hybrid screens; these binding partners now have been validated by co-immunoprecipitation experiments. Further functional characterization of interactions between Setdb2 and the identified binding partners, which include an unannotated protein and other biologically interesting proteins, has been conducted. We have generated a conditional knockout mouse model lacking Setdb2 located on a frequently deleted chromosomal region. Phenotypic characterization of these knockouts on various mouse strain backgrounds has begun. The knockout mice could comprise a new model for chronic lymphocytic leukemia, for asthma, and/or for other cancers or human diseases. Specific antibodies against mouse and human Setdb2 have been developed. We have expressed recombinant Setdb2 in baculovirus to facilitate characterization of its biochemical activities.

我们对候选组蛋白甲基转移酶Setdb2在人和小鼠组织中的转录进行了全面分析，揭示了选择性剪接和嵌合转录物的组织特异性表达。我们在框架内发现了将Setdb2与相邻的独立基因Phf11连接起来的融合转录本，该基因编码一个含锌指的植物同源结构域基序。我们之前使用酵母两种杂交筛选方法发现了几种假定的组蛋白甲基转移酶的蛋白质结合伙伴；这些结合伙伴现在已经通过共免疫沉淀实验得到了验证。Setdb2与鉴定的结合伙伴（包括一个未注释的蛋白和其他生物学上感兴趣的蛋白）之间相互作用的进一步功能表征已经进行。我们已经建立了一个条件敲除小鼠模型，该模型缺乏位于经常被删除的染色体区域的Setdb2。这些基因敲除在不同小鼠品系背景下的表型表征已经开始。基因敲除小鼠可构成慢性淋巴细胞白血病、哮喘和/或其他癌症或人类疾病的新模型。针对小鼠和人类Setdb2的特异性抗体已经开发出来。我们在杆状病毒中表达了重组Setdb2，以方便表征其生化活性。