3 UTR Control of TRA-2 mRNA in C. elegans

3 线虫中 TRA-2 mRNA 的 UTR 控制

基本信息

批准号：
6441174
负责人：
ELIZABETH B. GOODWIN
金额：
$ 34.08万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
1995
资助国家：
美国
起止时间：
1995-01-01 至 2006-02-28
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): How are some genes that specify cell fates regulated by elements in their 3' untranslated region (3'UTR) so that cells adopt a given fate at the right time and place in development? We will investigate this fundamental question of biology by analyzing how the 3'UTR of the C. elegans sex determination gene tra-2 controls development. Two regulatory processes act via the 3'UTR. The first is crucial for hermaphrodite development, and controls translation of the tra-2 mRNA by two 3'UTR elements (called TGEs). GLD-l is a TGE dependent translational repressor, and the gene laf-1 is also required for control. The second regulation is essential for development of both hermaphrodites and males, and controls the activity of the zinc finger transcription factor, TRA-1. TRA-1 binding to the tra-2 3'UTR inhibits TRA-1 transcriptional regulatory activity by promoting the nuclear export of the protein. The export of tra-2 mRNA and TRA-1 are mutually dependent on one another suggesting they exit the nucleus as a complex. In the next five years, we propose to investigate how the tra-2 3'UTR controls development. First using transgenic and biochemical assays, we will investigate the molecular mechanism by which TGEs and GLD-1 inhibit translation. Recent evidence suggests that GLD-1 inhibits translation via the Poly(A) Binding Protein (PAB). We will generate mutations in GLD-1 that disrupts the GLD-1/PAB interaction, and ask if these proteins fail to repress translation. In addition, we will ask if TRA-1 also affects TGE dependent control. Second, we will investigate if laf-1 is a regulatory RNA, as molecular analysis suggests the gene encodes no significant open reading frames. Also, we will ask when and where laf-1 gene product is expressed. Third, we will investigate how regulated nuclear export of tra-2 mRNA and TRA-1 is accomplished. Preliminary data indicates CeNXF-2, a protein similar to Mex67/TAP, is necessary for tra-2 mRNA nuclear retention. Using transgenes and RNA gel shift, we will confirm and extend these findings. In addition, we will identify the sequences of TRA-1 and the tra-2 3'UTR required for TRA-1 RNA binding and export. Finally, we will investigate the role of export in sex determination by asking if any known sex determining genes control TRA-1/tra-2 mRNA nuclear export. The health relatedness of this work derives from its contribution to an understanding of fundamental control of gene activity by 3'UTRs. The TGE control is present in several metazoans, including man, and governs the translation of the human oncogene GLI1, suggesting this control plays a role in suppressing tumorigenesis.

描述（由申请人提供）：某些指定细胞命运的基因如何由3'未翻译区域（3'UTR）中的元素调节，以便细胞在正确的时间和地点采用给定的命运？我们将通过分析3'UTR如何调查生物学的基本问题秀丽隐杆线虫性确定基因Tra-2控制着发展。二监管过程通过3'UTR。第一个对雌雄同体至关重要开发，并通过两个3'UTR元素控制Tra-2 mRNA的翻译（称为tges）。 GLD-L是TGE依赖性翻译阻遏物，并且基因控制也需要LAF-1。第二个法规对于雌雄同体和男性的发展，并控制锌指转录因子，tra-1。 Tra-1与Tra-2 3'utr结合通过促进核抑制Tra-1的转录调节活性蛋白质的出口。 Tra-2 mRNA和TRA-1的导出是相互的彼此取决于表明它们将核作为复合物退出。在接下来的五年，我们建议研究Tra-2 3'utr如何控制发展。首先使用转基因和生化测定法，我们将研究 TGES和GLD-1抑制翻译的分子机制。最近的证据表明，GLD-1通过poly（a）结合抑制翻译蛋白质（PAB）。我们将在GLD-1中产生突变，以破坏GLD-1/PAB 相互作用，并询问这些蛋白质是否无法抑制翻译。在此外，我们将询问TRA-1是否还会影响TGE依赖控制。第二，我们正如分子分析所表明的那样，将研究LAF-1是否是调节性RNA 该基因没有编码明显的开放式阅读框架。另外，我们将询问何时和表达LAF-1基因产物的位置。第三，我们将调查如何受到监管完成TRA-2 mRNA和TRA-1的核出口。初步数据表明CENXF-2（一种类似于MEX67/TAP的蛋白质）对于Tra-2 mRNA是必需的核保留。使用转基因和RNA凝胶转移，我们将确认并扩展这些发现。此外，我们将确定Tra-1和 TRA-2 3'UTR需要Tra-1 RNA结合和导出。最后，我们会的通过询问是否有任何已知性别来调查出口在性别确定中的作用确定基因控制TRA-1/TRA-2 mRNA核输出。健康这项工作的相关性来自其对理解的贡献 3'UTRS对基因活性的基本控制。 TGE控制存在于包括人在内的几个后生动物，并控制着人类的翻译 Oncogene Gli1，表明此控制在抑制中起作用肿瘤发生。