权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

The role of nuclear pore component Megator in gene expression and dosage compensation

核孔成分Megator在基因表达和剂量补偿中的作用

基本信息

批准号：
9760952
负责人：
JENNIFER ALEMAN
金额：
$ 4.5万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-01 至 2021-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9760952
关键词：
Animals Binding Biochemical Biogenesis Biological Assay Carrier Proteins Cell Nucleus Cells ChIP-seq Chromatin Chromosomes Complex Data Development Developmental Process Disease Dosage Compensation (Genetics)Drosophila genus Drosophila melanogaster Ensure Epigenetic Process Exhibits Face Female Gene Dosage Gene Expression Gene Expression Regulation Genes Genetic Genetic Transcription Genome Goals Homologous Gene Image Immunofluorescence Immunologic Investigation Knowledge Link Male Sterility Mediating Messenger RNA Mitotic Molecular Molecular Structure Mutation Nature Nuclear Nuclear Envelope Nuclear Matrix Nuclear Pore Nuclear Pore Complex Nuclear Pore Complex Proteins Organism Pattern Process RNA RNA Polymerase II RNA Processing RNA Splicing RNA analysis Regulation Regulator Genes Reporting Role Salivary Glands Stains Structure Testing Tissues Transcriptional Regulation Untranslated RNA Up-Regulation Work X Chromosome Yeasts autosome dosage exosome experimental study female fertility insight mRNA Export mRNA Surveillance male novel promoter protein transport scaffold sex single molecule transcriptome sequencing

项目摘要

Project Summary The goal of this proposal is to determine the role of the nuclear pore complex (NPC) protein Megator (Mtor) in regulation of gene expression by determining the mechanism via which Mtor influences the epigenetic phenomenon of dosage compensation in Drosophila melanogaster. Mtor makes up the nuclear basket of the NPC and appears to form an intranuclear matrix-like structure that, binds along chromatin in post-mitotic, polytenized salivary gland nuclei. Since Mtor has been implicated in regulation of RNA biogenesis processes in yeast and in the process of dosage compensation in Drosophila, I examined the effect of Mtor on localization of a non-coding RNA that is part of the dosage compensation (MSL) complex, roX1. The Drosophila MSL complex functions to upregulate expression of the single male X chromosome two-fold. Using single molecule RNA FISH in larval salivary gland nuclei, I detected an increase in nuclear soluble roX1 that is not associated with the X chromosome in addition to normal roX1 targeting in Mtor-depleted conditions. The increased presence of nuclear soluble roX1 was found to be due to increased levels of transcription of roX1 in male nuclei. In addition to rox1, a number of other X-linked genes exhibited a male-specific increase in expression upon depletion of Mtor, suggesting that normally Mtor functions to restrict dosage compensated gene expression. Aim 1 will test the hypothesis that Mtor restricts gene expression on the male X chromosome. RNA- Seq will be performed on larval salivary glands from both control and Mtor depleted conditions as well as from both males and females. In addition to examining gene expression changes, I will also be able to examine RNA processing differences by RNA-Seq and RNA FISH in order to begin investigation into the mechanism of Mtor’s effects on gene expression. Aim 2 will test the hypothesis that Mtor’s effects on gene expression are dependent on an interaction with components of the MSL complex and of the nuclear exosome, using assays for chromatin binding and for genetic interactions. These studies will uncover a novel gene regulatory role for a nuclear-scaffold forming nucleoporin in the context of dosage compensation. These aims will help to elucidate a role for Mtor in RNA biogenesis in the context of a developing organism as well as expand our understanding of the NPC’s role in gene regulation.

项目总结