Genetic Innovation of Cytoskeletal Proteins for Specialized Functions in the Male Germline

男性种系中具有特殊功能的细胞骨架蛋白的遗传创新

基本信息

批准号：
10534251
负责人：
Courtney Megan Schroeder
金额：
$ 24.9万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10534251
关键词：
Acceleration Actin-Binding Protein Actins Amino Acid Substitution Animals Antibodies Automobile Driving Biochemistry Biological Assay Biological Process Candidate Disease Gene Cell division Cytoskeletal Proteins Cytoskeleton Defect Drosophila genome Drosophila genus Drosophila inturned protein Eukaryota Event Evolution Exhibits Family Family member Fertility Future Gene Duplication Gene Family Gene Proteins Generations Genes Genetic Histologic Histology Infertility Insecta Interdisciplinary Study Knock-out Knockout Mice Male Infertility Mammals Men&apos s Role Microscopy Microtubules Modeling Molecular Mus Nuclear Orthologous Gene Phenotype Play Postdoctoral Fellow Protein Biochemistry Protein Family Proteins Recording of previous events Recurrence Reproduction Research Role Spermatogenesis Structure Surveys System Testing Testis Training Work cell motility comparative genomics cost design fitness fitness test fly genetic signature innovation male male fertility mammalian genome member novel paralogous gene pressure programs recruit reproductive reproductive fitness skills sperm cell tool

项目摘要

PROJECT SUMMARY Cytoskeletal proteins are fundamental in many biological processes among all eukaryotes. Given their essentiality, they are conventionally thought to be highly conserved. However, signatures of genetic innovation in the form of gene-family expansions and accelerated amino-acid substitutions are found among key cytoskeletal family members. Rapid molecular divergence between closely related species is unexpected in conserved protein families and is indicative of an adaptive advantage for sequence innovation. This proposal will investigate the causes and consequences of this dramatic diversification of cytoskeletal proteins. Actin- related proteins (Arps) are one cytoskeletal family that has ancient evolutionary origins yet exhibits recurrent genetic innovation in both mammals and insects. Unlike the well-conserved members of the Arp superfamily, divergent Arps are testis-specific in expression, suggesting they have acquired specialized roles for male reproduction. Yet the biological process that drives their innovation is unknown. The localization of gametic Arps in Drosophila suggests they play roles at germline-specific actin structures. This proposal will address the hypothesis that gametic Arps facilitate the generation of actin structures that are critical for male reproductive fitness. Aim 1 will uncover the function of one gametic Arp found in all Drosophila species, Arp53D, which localizes to two germline actin structures during late spermatogenesis. D. melanogaster, which has many well- established genetic tools and only one gametic Arp, provides an advantageous system in which to investigate the function of a divergent Arp at actin and probe how its structural diversification allows for novel roles. Aim 2 will extend the analysis to mammalian gametic Arps to gain a deeper understanding of the potentially shared biological processes that are driving Arp innovation among multiple phyla. A few pieces of evidence suggest that mammalian gametic Arps play actin regulatory roles, much like Arp53D. Aim 2 will take a comprehensive approach to elucidate their evolutionary histories and localization with respect to actin, while characterizing the male infertility phenotype of one Arp with an established mouse knockout model. Finally, preliminary findings suggest additional cases of rapidly evolving cytoskeletal proteins, beyond Arps, are present in the male germline. Aim 3 will systematically search Drosophila and mammalian genomes for rapid evolution and gene duplications among cytoskeletal proteins that are testis-enriched in expression. This aim will yield additional cases of genetic innovation that will serve as projects in the applicant’s future lab to broadly understand the adaptive sequence plasticity of cytoskeletal proteins and what drives their specialization for male reproduction. To launch this project, the applicant requires training in fertility assays, mouse testis histology and additional evolutionary analyses. She will ultimately use her background in biochemistry to study diversifying cytoskeletal proteins at a mechanistic level.

项目概要细胞骨架蛋白是所有真核生物许多生物过程的基础。本质上，它们通常被认为是高度保守的，然而，基因创新的特征。以基因家族扩展和加速氨基酸取代的形式发现了关键的细胞骨架家族成员之间密切相关的物种之间的快速分子分化是出乎意料的。保守的蛋白质家族，表明该提议具有适应性优势。将研究细胞骨架蛋白这种戏剧性多样化的原因和后果。相关蛋白 (Arps) 是一种细胞骨架家族，具有古老的进化起源，但表现出反复出现的特征与 Arp 超家族中保存完好的成员不同，哺乳动物和昆虫的基因创新。不同的 Arps 在表达上具有睾丸特异性，这表明它们已经获得了男性的特殊作用然而，驱动它们创新的生物过程尚不清楚。果蝇中的 Arps 表明它们在种系特异性肌动蛋白结构中发挥作用。配子 Arps 促进对雄性繁殖至关重要的肌动蛋白结构的生成的假设目标 1 将揭示所有果蝇物种中发现的一种配子 Arp（Arp53D）的功能。在精子发生后期定位于两个种系肌动蛋白结构，其具有许多良好的结构。已建立的遗传工具和只有一个配子 Arp，提供了一个有利的系统来研究肌动蛋白上不同的 Arp 的功能，并探讨其结构多样化如何发挥新的作用。将把分析扩展到哺乳动物配子 Arps，以更深入地了解潜在的共享一些证据表明，推动 Arp 在多个门中创新的生物过程。哺乳动物配子 Arps 发挥肌动蛋白调节作用，就像 Arp53D 一样，Aim 2 将发挥全面作用。阐明其关于肌动蛋白的进化历史和定位的方法，同时表征最后，初步发现了一种 Arp 的雄性不育表型与已建立的小鼠敲除模型。表明除了 Arps 之外，男性体内还存在快速进化的细胞骨架蛋白的其他情况 Aim 3 将系统地搜索果蝇和哺乳动物基因组的快速进化和基因。睾丸表达丰富的细胞骨架蛋白之间的重复这一目标将产生额外的结果。基因创新案例将作为申请人未来实验室的项目，以广泛了解细胞骨架蛋白的适应性序列可塑性以及驱动其男性生殖专业化的因素。为了启动该项目，申请人需要接受生育力测定、小鼠睾丸组织学和其他方面的培训她最终将利用她的生物化学背景来研究细胞骨架的多样化。机械水平上的蛋白质。