Cell Biological mechanisms of centromere drive

着丝粒驱动的细胞生物学机制

• 批准号: 10605289
• 负责人: Michael Lampson
• 金额: $ 42.66万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10605289
• 关键词: Address; Biological; Biological Models; Cell division; Cells; Centromere; Chemicals; Chromosome Segregation; Chromosomes; Conflict (Psychology); DNA; DNA Sequence; Defect; Development; Ensure; Evolution; Female; Genetic; Genetic Materials; Genome; Goals; Hybrids; Inherited; Lead; Link; Meiosis; Microtubules; Modeling; Molecular; Molecular Abnormality; Molecular Evolution; Mus; Pathway interactions; Post-Translational Protein Processing; Pregnancy loss; Process; Proteins; Recurrence; Regulation; Reproductive Biology; Satellite DNA; System; Testing; Tubulin; Variant; Work; cost; egg; fitness; mouse model; optogenetics; recruit; reproductive fitness; segregation; sperm cell; theories; transmission process; zygote

The seemingly straightforward function of the centromere in directing chromosome segregation is difficult to reconcile with multiple complexities of the underlying molecular machinery, particularly rapid evolution of both centromere DNA and proteins and seemingly redundant pathways linking the DNA to spindle microtubules. This project focuses on centromere drive as a key to unlocking centromere complexity. Selfish centromere DNA sequences bias their transmission to the egg in female meiosis, while centromere proteins evolve to suppress fitness costs of drive while maintaining essential centromere functions. Our recent work determined how selfish centromeres interact with spindle microtubules to bias their segregation. We developed mouse model systems exploiting natural variation in mouse centromere DNA, defined tubulin detyrosination as the key post-translational modification creating meiotic spindle asymmetry, showed that microtubule-destabilizing proteins act as drive effectors exploited by selfish centromeres, established an integrated model for both drive and suppression, and sequenced Murinae genomes for molecular evolution analyses to identify rapidly evolving centromere proteins. Our progress represents crucial steps towards understanding the centromere drive conflict but leaves key gaps in our understanding of drive and suppression and centromere protein evolution, which are addressed in this proposal. First, we will determine how selfish centromeres interact with an asymmetric spindle to bias their segregation. Our previous findings suggest a hypothesis that we will test by manipulating microtubule destabilizing activities at centromeres in live cells, using chemical optogenetic approaches that we developed. Second, we will test whether genetically different centromeres differentially recruit centromere proteins, a central but untested component of the centromere drive theory. Using hybrid mouse zygotes with divergent maternal and paternal centromere satellite DNA sequences as a model system, we will determine if rapidly evolving centromere protein interact differentially with different centromere DNA sequences. Third, we will test for reproductive fitness costs associated with functional differences between centromeres, taking advantage of our hybrid mouse model systems in which paired homologous chromosomes in meiosis have divergent centromeres. Fourth, we will test the concept that centromere proteins have evolved to suppress costs due to functional differences between centromeres, which has been the most challenging part of the drive theory to address experimentally. With tractable experimental systems, a mechanistic model for drive and suppression, and molecular evolution analyses of centromere proteins in place, we will address this challenge by testing whether recurrent changes in rapidly evolving centromere proteins have functional implications consistent with our model. Overall, by investigating centromeres in the context of genetic conflict, this project represents a unique contribution to studies of chromosome segregation and inheritance, with broad consequences for reproductive biology and chromosome evolution.

着丝粒在指导染色体分离方面的功能看似简单，但很难理解。 与潜在分子机制的多重复杂性相协调，特别是两者的快速进化 着丝粒 DNA 和蛋白质以及连接 DNA 和纺锤体微管的看似多余的途径。 该项目重点关注着丝粒驱动作为解锁着丝粒复杂性的关键。自私的着丝粒 DNA 序列在雌性减数分裂中偏向于向卵子的传输，而着丝粒蛋白则进化为 抑制驱动力的健康成本，同时维持基本的着丝粒功能。我们近期的工作确定 自私的着丝粒如何与纺锤体微管相互作用以偏向其分离。我们开发了鼠标 利用小鼠着丝粒 DNA 自然变异的模型系统，将微管蛋白脱酪氨酸定义为关键 翻译后修饰产生减数分裂纺锤体不对称性，表明微管不稳定 蛋白质充当自私着丝粒利用的驱动效应器，建立了两种驱动的集成模型 和抑制，并对鼠亚科基因组进行测序，进行分子进化分析，以快速识别 不断进化的着丝粒蛋白。我们的进展代表着理解着丝粒的关键步骤 驱动冲突，但在我们对驱动和抑制以及着丝粒蛋白的理解中留下了关键空白 进化，这在本提案中得到解决。首先，我们将确定自私的着丝粒如何与 一个不对称的纺锤体来偏向它们的分离。我们之前的发现提出了一个假设，我们将通过以下方式进行检验 使用化学光遗传学操纵活细胞着丝粒的微管不稳定活动 我们开发的方法。其次，我们将测试遗传不同的着丝粒是否存在差异 招募着丝粒蛋白，这是着丝粒​​驱动理论的核心但未经测试的组成部分。使用混合动力 具有不同母本和父本着丝粒卫星DNA序列的小鼠受精卵作为模型系统， 我们将确定快速进化的着丝粒蛋白是否与不同的着丝粒 DNA 存在差异性相互作用 序列。第三，我们将测试与功能差异相关的生殖健康成本 着丝粒，利用我们的杂交小鼠模型系统，其中配对同源染色体 减数分裂时有不同的着丝粒。第四，我们将检验着丝粒蛋白进化的概念 抑制由于着丝粒之间的功能差异而产生的成本，这是最具挑战性的 驱动理论的一部分需要通过实验来解决。通过易于处理的实验系统，机械模型 对于驱动和抑制以及着丝粒蛋白的分子进化分析，我们将解决 通过测试快速进化的着丝粒蛋白的反复变化是否具有功能性来应对这一挑战 影响与我们的模型一致。总的来说，通过研究遗传冲突背景下的着丝粒， 该项目对染色体分离和遗传的研究做出了独特的贡献，具有广泛的意义 对生殖生物学和染色体进化的影响。

项目成果

Chromosome instability in tumor cells due to defects in Aurora B mediated error correction at kinetochores.

• DOI: 10.1080/15384101.2018.1553340
• 发表时间: 2018
• 期刊: Cell cycle (Georgetown, Tex.)
• 作者: Huang H;Lampson M;Efimov A;Yen TJ
• 通讯作者: Yen TJ

Parallel pathways for recruiting effector proteins determine centromere drive and suppression.

• DOI: 10.1016/j.cell.2021.07.037
• 发表时间: 2021-09-16
• 期刊: Cell
• 影响因子: 64.5
• 作者: Kumon T;Ma J;Akins RB;Stefanik D;Nordgren CE;Kim J;Levine MT;Lampson MA
• 通讯作者: Lampson MA

Evolution of eukaryotic centromeres by drive and suppression of selfish genetic elements.

• DOI: 10.1016/j.semcdb.2022.03.026
• 发表时间: 2022-08
• 期刊: SEMINARS IN CELL & DEVELOPMENTAL BIOLOGY
• 影响因子: 7.3
• 作者: Kumon, Tomohiro;Lampson, Michael A.
• 通讯作者: Lampson, Michael A.

Reversible optogenetic control of protein function and localization.

蛋白质功能和定位的可逆光遗传学控制。

• DOI: 10.1016/bs.mie.2019.05.002
• 发表时间: 2019
• 期刊: Methods in enzymology
• 作者: Wu,DanielZ;Lackner,RachelM;Aonbangkhen,Chanat;Lampson,MichaelA;Chenoweth,DavidM
• 通讯作者: Chenoweth,DavidM

Spindle asymmetry drives non-Mendelian chromosome segregation.

• DOI: 10.1126/science.aan0092
• 发表时间: 2017-11-03
• 期刊: Science (New York, N.Y.)
• 作者: Akera T;Chmátal L;Trimm E;Yang K;Aonbangkhen C;Chenoweth DM;Janke C;Schultz RM;Lampson MA
• 通讯作者: Lampson MA