LOCALIZATION OF MYOSIN MRNA BY 3'UTR SEQUENCES

通过 3UTR 序列对肌球蛋白 mRNA 进行定位

• 批准号: 2415513
• 负责人: WILLARD W SHARP
• 金额: $ 2.86万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-05-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2415513
• 关键词: RNA binding protein; antisense nucleic acid; fusion gene; gel mobility shift assay; heart cell; heart contraction; messenger RNA; muscle cells; myosins; nucleic acid sequence; protein biosynthesis; tissue /cell culture; ultraviolet radiation

Synthesis of myosin isoforms and their incorporation into myofibrils affects cardiac function. The goal of the proposed research is to answer the following hypotheses: I)The 3'UTR of alpha and beta myosin mRNA regulates their intracellular localization during changes in contractile activity 2)Binding of mRNPs to the 3'UTR is regulated by contractile activity. To test the function of the 3'UTR, it is necessary to make constructs that isolate this mRNA sequence from the rest of the myosin mRNA. This requires fusion constructs that attach the hypothetical regulatory 3'UTR to sequences coding for a reporter protein. Behavior of the reporter mRNA and translation of the reporter protein in transfected myocytes then give information about the function of the myosin 3'UTR. 3'UTR regulation of mRNA localization is likely to be mediated by mRNA binding proteins. Gel retardation assays coupled with UV crosslinking assays will be used to investigate mRNP binding to 3'UTR sequences. The use of antisense oligonuclotides will determine which sequences are important for mRNA localization and mRNP binding.

肌球蛋白同工型的合成及其掺入肌原纤维 影响心脏功能。拟议研究的目的是 回答以下假设： i）Alpha和Beta肌球蛋白mRNA的3'Utr调节他们 收缩活动变化期间的细胞内定位 2）MRNP与3'UTR的结合受收缩活性调节。 要测试3'UTR的功能，有必要制作构造 将这种mRNA序列与其他肌球蛋白mRNA分离出来。这 需要附加假设调节的融合结构 3'UTR用于编码报告基因蛋白的序列。行为 记者mRNA和转染中报告蛋白的翻译 然后，肌细胞提供有关肌球蛋白3'UTR功能的信息。 mRNA定位的3'UTR调节可能是由mRNA介导的 结合蛋白。凝胶延迟测定与紫外线交联 测定将用于研究与3'UTR序列的MRNP结合。 反义寡核氯肽的使用将确定哪些序列 对于mRNA定位和MRNP结合非常重要。