mRNP granule proteins Fmrp and Dcp1a differentially regulate mRNP complexes to contribute to control of muscle stem cell quiescence and activation.

IF 8.3 2区 材料科学 Q1 MATERIALS SCIENCE, MULTIDISCIPLINARY
Nainita Roy, Swetha Sundar, Malini Pillai, Farah Patell-Socha, Sravya Ganesh, Ajoy Aloysius, Mohammed Rumman, Hardik Gala, Simon M Hughes, Peter S Zammit, Jyotsna Dhawan
{"title":"mRNP granule proteins Fmrp and Dcp1a differentially regulate mRNP complexes to contribute to control of muscle stem cell quiescence and activation.","authors":"Nainita Roy,&nbsp;Swetha Sundar,&nbsp;Malini Pillai,&nbsp;Farah Patell-Socha,&nbsp;Sravya Ganesh,&nbsp;Ajoy Aloysius,&nbsp;Mohammed Rumman,&nbsp;Hardik Gala,&nbsp;Simon M Hughes,&nbsp;Peter S Zammit,&nbsp;Jyotsna Dhawan","doi":"10.1186/s13395-021-00270-9","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>During skeletal muscle regeneration, satellite stem cells use distinct pathways to repair damaged myofibers or to self-renew by returning to quiescence. Cellular/mitotic quiescence employs mechanisms that promote a poised or primed state, including altered RNA turnover and translational repression. Here, we investigate the role of mRNP granule proteins Fragile X Mental Retardation Protein (Fmrp) and Decapping protein 1a (Dcp1a) in muscle stem cell quiescence and differentiation.</p><p><strong>Methods: </strong>Using isolated single muscle fibers from adult mice, we established differential enrichment of mRNP granule proteins including Fmrp and Dcp1a in muscle stem cells vs. myofibers. We investigated muscle tissue homeostasis in adult Fmr1-/- mice, analyzing myofiber cross-sectional area in vivo and satellite cell proliferation ex vivo. We explored the molecular mechanisms of Dcp1a and Fmrp function in quiescence, proliferation and differentiation in a C2C12 culture model. Here, we used polysome profiling, imaging and RNA/protein expression analysis to establish the abundance and assembly status of mRNP granule proteins in different cellular states, and the phenotype of knockdown cells.</p><p><strong>Results: </strong>Quiescent muscle satellite cells are enriched for puncta containing the translational repressor Fmrp, but not the mRNA decay factor Dcp1a. MuSC isolated from Fmr1<sup>-/-</sup> mice exhibit defective proliferation, and mature myofibers show reduced cross-sectional area, suggesting a role for Fmrp in muscle homeostasis. Expression and organization of Fmrp and Dcp1a varies during primary MuSC activation on myofibers, with Fmrp puncta prominent in quiescence, but Dcp1a puncta appearing during activation/proliferation. This reciprocal expression of Fmrp and Dcp1a puncta is recapitulated in a C2C12 culture model of quiescence and activation: consistent with its role as a translational repressor, Fmrp is enriched in non-translating mRNP complexes abundant in quiescent myoblasts; Dcp1a puncta are lost in quiescence, suggesting stabilized and repressed transcripts. The function of each protein differs during proliferation; whereas Fmrp knockdown led to decreased proliferation and lower cyclin expression, Dcp1a knockdown led to increased cell proliferation and higher cyclin expression. However, knockdown of either Fmrp or Dcp1a led to compromised differentiation. We also observed cross-regulation of decay versus storage mRNP granules; knockdown of Fmrp enhances accumulation of Dcp1a puncta, whereas knockdown of Dcp1a leads to increased Fmrp in puncta.</p><p><strong>Conclusions: </strong>Taken together, our results provide evidence that the balance of mRNA turnover versus utilization is specific for distinct cellular states.</p>","PeriodicalId":5,"journal":{"name":"ACS Applied Materials & Interfaces","volume":" ","pages":"18"},"PeriodicalIF":8.3000,"publicationDate":"2021-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s13395-021-00270-9","citationCount":"1","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"ACS Applied Materials & Interfaces","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1186/s13395-021-00270-9","RegionNum":2,"RegionCategory":"材料科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"MATERIALS SCIENCE, MULTIDISCIPLINARY","Score":null,"Total":0}
引用次数: 1

Abstract

Background: During skeletal muscle regeneration, satellite stem cells use distinct pathways to repair damaged myofibers or to self-renew by returning to quiescence. Cellular/mitotic quiescence employs mechanisms that promote a poised or primed state, including altered RNA turnover and translational repression. Here, we investigate the role of mRNP granule proteins Fragile X Mental Retardation Protein (Fmrp) and Decapping protein 1a (Dcp1a) in muscle stem cell quiescence and differentiation.

Methods: Using isolated single muscle fibers from adult mice, we established differential enrichment of mRNP granule proteins including Fmrp and Dcp1a in muscle stem cells vs. myofibers. We investigated muscle tissue homeostasis in adult Fmr1-/- mice, analyzing myofiber cross-sectional area in vivo and satellite cell proliferation ex vivo. We explored the molecular mechanisms of Dcp1a and Fmrp function in quiescence, proliferation and differentiation in a C2C12 culture model. Here, we used polysome profiling, imaging and RNA/protein expression analysis to establish the abundance and assembly status of mRNP granule proteins in different cellular states, and the phenotype of knockdown cells.

Results: Quiescent muscle satellite cells are enriched for puncta containing the translational repressor Fmrp, but not the mRNA decay factor Dcp1a. MuSC isolated from Fmr1-/- mice exhibit defective proliferation, and mature myofibers show reduced cross-sectional area, suggesting a role for Fmrp in muscle homeostasis. Expression and organization of Fmrp and Dcp1a varies during primary MuSC activation on myofibers, with Fmrp puncta prominent in quiescence, but Dcp1a puncta appearing during activation/proliferation. This reciprocal expression of Fmrp and Dcp1a puncta is recapitulated in a C2C12 culture model of quiescence and activation: consistent with its role as a translational repressor, Fmrp is enriched in non-translating mRNP complexes abundant in quiescent myoblasts; Dcp1a puncta are lost in quiescence, suggesting stabilized and repressed transcripts. The function of each protein differs during proliferation; whereas Fmrp knockdown led to decreased proliferation and lower cyclin expression, Dcp1a knockdown led to increased cell proliferation and higher cyclin expression. However, knockdown of either Fmrp or Dcp1a led to compromised differentiation. We also observed cross-regulation of decay versus storage mRNP granules; knockdown of Fmrp enhances accumulation of Dcp1a puncta, whereas knockdown of Dcp1a leads to increased Fmrp in puncta.

Conclusions: Taken together, our results provide evidence that the balance of mRNA turnover versus utilization is specific for distinct cellular states.

Abstract Image

Abstract Image

Abstract Image

mRNP颗粒蛋白Fmrp和Dcp1a对mRNP复合物的差异调控有助于控制肌肉干细胞的静止和激活。
背景:在骨骼肌再生过程中,卫星干细胞通过不同的途径修复受损的肌纤维或通过恢复静止状态进行自我更新。细胞/有丝分裂静止采用促进平衡或启动状态的机制,包括改变的RNA周转和翻译抑制。在这里,我们研究了mRNP颗粒蛋白脆性X智力迟钝蛋白(Fmrp)和脱帽蛋白1a (Dcp1a)在肌肉干细胞静止和分化中的作用。方法:利用分离的成年小鼠单个肌纤维,我们建立了肌肉干细胞与肌纤维中mRNP颗粒蛋白(包括Fmrp和Dcp1a)的差异富集。我们研究了成年Fmr1-/-小鼠的肌肉组织稳态,分析了体内肌纤维横截面积和体外卫星细胞增殖。我们在C2C12培养模型中探讨了Dcp1a和Fmrp功能在静止、增殖和分化中的分子机制。在这里,我们使用多聚体分析、成像和RNA/蛋白表达分析来建立mRNP颗粒蛋白在不同细胞状态下的丰度和组装状态,以及敲低细胞的表型。结果:静止肌卫星细胞中含有翻译抑制因子Fmrp,但不含mRNA衰变因子Dcp1a。从Fmr1-/-小鼠中分离的MuSC表现出增殖缺陷,成熟肌纤维的横截面积减少,表明Fmrp在肌肉稳态中的作用。肌纤维上Fmrp和Dcp1a的表达和组织在MuSC的原发性激活过程中存在差异,Fmrp点在静止状态突出,而Dcp1a点在激活/增殖过程中出现。这种Fmrp和Dcp1a点的相互表达在C2C12的静止和激活培养模型中得到了概括:与Fmrp作为翻译抑制因子的作用一致,Fmrp在静止的成肌细胞中丰富的非翻译mRNP复合物中富集;Dcp1a点在静止状态下丢失,表明转录本稳定和抑制。每种蛋白在增殖过程中的功能是不同的;Fmrp敲低导致细胞增殖减少和细胞周期蛋白表达降低,而Dcp1a敲低导致细胞增殖增加和细胞周期蛋白表达升高。然而,Fmrp或Dcp1a的敲低都会导致分化受损。我们还观察到mRNP颗粒的衰变与储存的交叉调节;敲低Fmrp可增强Dcp1a点的积累,而敲低Dcp1a可导致点中Fmrp的增加。结论:综上所述,我们的研究结果提供了证据,证明mRNA周转与利用的平衡是特定于不同细胞状态的。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
ACS Applied Materials & Interfaces
ACS Applied Materials & Interfaces 工程技术-材料科学:综合
CiteScore
16.00
自引率
6.30%
发文量
4978
审稿时长
1.8 months
期刊介绍: ACS Applied Materials & Interfaces is a leading interdisciplinary journal that brings together chemists, engineers, physicists, and biologists to explore the development and utilization of newly-discovered materials and interfacial processes for specific applications. Our journal has experienced remarkable growth since its establishment in 2009, both in terms of the number of articles published and the impact of the research showcased. We are proud to foster a truly global community, with the majority of published articles originating from outside the United States, reflecting the rapid growth of applied research worldwide.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信