Mechanism and effects of communication between actin and gene regulatory complexes

肌动蛋白与基因调控复合物之间通讯的机制和影响

基本信息

批准号：
10653689
负责人：
Ryan David Mohan
金额：
$ 38.56万
依托单位：
UNIVERSITY OF MISSOURI KANSAS CITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10653689
关键词：
Actins Affinity Chromatography Antibodies Binding Biological Process Blindness Cell Nucleus Cells Characteristics Column Chromatography Communication Complex Coupled Cytoplasm Cytoskeletal Modeling Cytoskeleton DNA Repair Pathway Defect Disease Dominant-Negative Mutation Ensure Enzymes Eye F-Actin Functional disorder Gene Expression Genetic Transcription Genome Homologous Protein Image Immunoprecipitation In Situ Hybridization Knowledge Lamins Lead Link Location Longevity Mass Spectrum Analysis Measures Membrane Messenger RNA Molecular Mutate Mutation Nerve Degeneration Nervous System Nuclear Nuclear Inner Membrane Nuclear Pore Nuclear Pore Complex Nuclear Protein Pathway interactions Phenotype Polymers Proteins Publishing Regulation Regulator Genes Regulatory Pathway Reporter Retina Role SAGA SCA7 protein Shapes Site Symptoms System Tail Testing Tissues Transcript Transcription Coactivator Visualization WAVE protein experimental study fly in vivo insight mRNA Export mutant novel novel therapeutics polyglutamine polymerization prevent promoter protein complex recruit repaired spatiotemporal ubiquitin isopeptidase

项目摘要

Abstract Although many studies have demonstrated correlations between cytoskeletal dynamics, genome organization, and gene expression, the underlying mechanisms linking them remain unclear. Recently, we discovered crosstalk between the Wiskott-Aldrich syndrome protein family verprolin homolog (WAVE) regulatory complex (WRC), which promotes actin polymerization, and the Spt-Ada-Gcn5-acetyltransferase (SAGA) complex, a transcriptional coactivator. Their relationship is established through the sharing of subunits comprising the SAGA deubiquitinase module, including the deubiquitinase Non-stop. Deubiquitinase module mutations—for example, polyglutamine expansion in ataxin 7 (ATXN7)—lead to a spectrum of phenotypes not explained by SAGA’s transcriptional coactivator function, including nervous system degeneration and blindness. In both the nervous system and the eye, the WRC is an essential promoter of actin polymerization, which is facilitated by the constitutively active enzymatic subunit WAVE. WAVE activity and localization are regulated by the remaining WRC subunits, ensuring spatial and temporal control of actin polymerization. Misregulation of actin polymerizing complexes results in a similar spectrum of phenotypes as seen in SAGA mutants, including nervous system degeneration and blindness. This suggests that SAGA is important in the nervous system and eye because it is required to control the WRC in these tissues. We found that the SAGA deubiquitinase module leaves SAGA to bind the WRC. There, Non-stop deubiquitinates WAVE, increasing its level in both the cytoplasm and the nucleus. Therefore, we hypothesize that SAGA controls WRC complex composition, amount, and location; and it is through these activities that SAGA accomplishes functions we had previously attributed to SAGA alone. This hypotheses will be investigated in three aims. First, we will identify and characterize nuclear WAVE-containing complexes through affinity purification and column chromatography coupled to mass spectrometry. Because WAVE activity is regulated in part by its localization, the locations of these complexes will be determined, and the effects of Atxn7 polyglutamine expansion on complex composition and localization will be tested. Second, interactions between the SAGA deubiquitinase module and WRC will be disrupted in cells and flies to determine which SAGA/WRC functions require them. Lastly, the effects of SAGA deubiquitinase-WRC interactions on blindness and neurodegeneration will be investigated in flies, by disrupting them and measuring phenotypes characteristic of Atxn7 polyglutamine expansion. These studies will provide novel insight on the causes of neurodegeneration and blindness, in addition to the links between transcriptional and cytoskeletal regulatory complexes.

抽象的尽管许多研究已经证明了细胞骨架动力学、基因组组织、最近，我们发现，连接它们的潜在机制仍不清楚。 Wiskott-Aldrich 综合征蛋白家族 verprolin 同源物 (WAVE) 调节复合物之间的串扰 (WRC)，促进肌动蛋白聚合，以及 Spt-Ada-Gcn5-乙酰转移酶 (SAGA) 复合物，它们的关系是通过共享组成的亚基建立的。 SAGA 去泛素酶模块，包括去泛素酶不间断突变。例如，ataxin 7 (ATXN7) 中的聚谷氨酰胺扩增——导致一系列无法解释的表型 SAGA的转录共激活子功能，包括神经系统退化和失明。神经系统和眼睛中，WRC 是肌动蛋白聚合的重要促进剂，其促进作用是组成型活性酶亚基 WAVE 的活性和定位受 WAVE 的调节。剩余的 WRC 亚基，确保肌动蛋白聚合的空间和时间控制。聚合复合物会产生与 SAGA 突变体相似的表型谱，包括神经系统退化和失明这表明SAGA在神经系统和失明中很重要。眼睛，因为它需要控制这些组织中的 WRC 我们发现 SAGA 去泛素酶模块。让 SAGA 与 WRC 结合，Non-stop 使 WAVE 去泛素化，增加其在两个组织中的水平。因此，我们发现 SAGA 控制 WRC 复合物的组成， SAGA 正是通过这些活动完成了我们之前的职能我们将通过三个目标来研究这一假设。通过亲和纯化和柱色谱表征含有核 WAVE 的复合物由于 WAVE 活性部分受到其定位的调节，因此将确定这些复合物，以及 Atxn7 聚谷氨酰胺扩增对复合物的影响其次，将测试 SAGA 去泛素酶模块之间的相互作用。 WRC 将在细胞和苍蝇中被破坏，以确定哪些 SAGA/WRC 功能需要它们。 SAGA 去泛素酶-WRC 相互作用对失明和神经变性的影响将在果蝇，通过破坏它们并测量 Atxn7 聚谷氨酰胺扩增的表型特征。除了链接之外，研究还将提供关于神经退行性变和失明原因的新见解转录和细胞骨架调节复合物之间。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Deubiquitinases in Neurodegeneration.

神经变性中的去泛素酶。

DOI：
发表时间：
2022-02-05
期刊：
影响因子：
6
作者：
Bello, Abudu I;Goswami, Rituparna;Brown, Shelby L;Costanzo, Kara;Shores, Taylor;Allan, Shefaa;Odah, Revan;Mohan, Ryan D
通讯作者：
Mohan, Ryan D

The Molecular Basis of Spinocerebellar Ataxia Type 7.

7 型脊髓小脑共济失调的分子基础。

DOI：
发表时间：
2022
期刊：
影响因子：
0
作者：
Goswami, Rituparna;Bello, Abudu I;Bean, Joe;Costanzo, Kara M;Omer, Bwaar;Cornelio;Odah, Revan;Ahluwalia, Amit;Allan, Shefaa K;Nguyen, Nghi;Shores, Taylor;Aziz, N Ahmad;Mohan, Ryan D
通讯作者：
Mohan, Ryan D