Moa1与CENP-C和Rec8的相互作用及其在裂殖酵母减数分裂中的功能

doi:10.16288/j.yczz.24-035

遗传 ›› 2024, Vol. 46 ›› Issue (8): 649-660.doi: 10.16288/j.yczz.24-035

Moa1与CENP-C和Rec8的相互作用及其在裂殖酵母减数分裂中的功能

闵羽¹^,²(), 倪子涵¹^,², 马玲玲¹^,², 渡边嘉典¹^,²()

1.江南大学未来食品科学中心，无锡 214122
2.江南大学生物工程学院，无锡 214122

收稿日期:2024-03-08 修回日期:2024-05-07 出版日期:2024-08-20 发布日期:2024-05-14
通讯作者: 渡边嘉典，教授，研究方向：染色体分离机制。E-mail: ywatanabe@jiangnan.edu.cn
作者简介:闵羽，硕士研究生，专业方向：生物与医药。E-mail: 1401577396@qq.com

Functional roles of the interaction of Moa1 with CENP-C and Rec8 in meiosis of Schizosaccharomyces pombe

Yu Min¹^,²(), Zihan Ni¹^,², Lingling Ma¹^,², Yoshinori Watanabe¹^,²()

1. Science Center for Future Foods, Jiangnan University, Wuxi 214122, China
2. School of Biotechnology, Jiangnan University, Wuxi 214122, China

Received:2024-03-08 Revised:2024-05-07 Published:2024-08-20 Online:2024-05-14

摘要/Abstract

摘要：

减数分裂特异性调控分子Moa1定位到着丝粒受到动粒蛋白CENP-C的调控，同时Moa1参与黏连蛋白Rec8介导的着丝粒区域姐妹染色单体的黏连。为了研究这些蛋白质之间的相互作用，本研究利用酵母双杂交实验(yeast two-hybrid assay)测定分析了Moa1和CENP-C、Rec8之间的相互作用，并通过在Moa1中定点突变鉴定了与CENP-C和Rec8相互作用所需的一些氨基酸残基。实验结果表明，Moa1和CENP-C的相互作用对于Moa1参与调节姐妹动粒的单极附着很重要。然而，双杂交实验中与Rec8相互作用所需的Moa1的S143和T150突变没有显示出Moa1或Rec8功能的显著缺陷。这表明氨基酸残基的突变可能不足以干扰体内Moa1和Rec8之间的相互作用，需要进一步的研究来确定Moa1和Rec8的相互作用域。本研究揭示了影响减数分裂同源染色体分离的Moa1氨基酸位点，为减数分裂的染色体分离机制提供更深入的理解。

关键词: 减数分裂, 黏连蛋白, 动粒, 单取向

Abstract:

The localization of the meiotic specific regulatory molecule Moa1 to the centromere is regulated by the kinetochore protein CENP-C, and participates in the cohesion of sister chromatids in the centromere region mediated by the cohesin Rec8. To examine the interaction of these proteins, we analyzed the interactions between Moa1 and Rec8, CENP-C by yeast two-hybrid assays and identified several amino acid residues in Moa1 required for the interaction with CENP-C and Rec8. The results revealed that the interaction between Moa1 and CENP-C is crucial for the Moa1 to participate in the regulation of monopolar attachment of sister kinetochores. However, mutation at S143 and T150 of Moa1, which are required for interaction with Rec8 in the two-hybrid assay, did not show significant defects. Mutations in amino acid residues may not be sufficient to interfere with the interaction between Moa1 and Rec8 in vivo. Further research is needed to determine the interaction domain between Moa1 and Rec8. This study revealed specific amino acid sites at which Moa1 affects the meiotic homologous chromosome segregation, providing a deeper understanding of the mechanism of meiotic chromosome segregation.

Key words: meiosis, cohesin, kinetochore, mono-orientation

闵羽, 倪子涵, 马玲玲, 渡边嘉典. Moa1与CENP-C和Rec8的相互作用及其在裂殖酵母减数分裂中的功能[J]. 遗传, 2024, 46(8): 649-660.

Yu Min, Zihan Ni, Lingling Ma, Yoshinori Watanabe. Functional roles of the interaction of Moa1 with CENP-C and Rec8 in meiosis of Schizosaccharomyces pombe[J]. Hereditas(Beijing), 2024, 46(8): 649-660.

图/表 6

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菌株名称	基因型
PW632	h⁺ pat1^-114 3pk-moa1
PW1	h⁺ pat1-114 natMX6-3pk-moa1
M1	h^- leu1 imr1L-GFP mes1-B44 rec8-2A<<Kan rec12::hyg
M2	h⁺ mes1-B44 rec8-2A<<Kan rec12::hyg
M3	h⁹⁰ mei4::hyg
PM11	h^- leu1 imr1L-GFP mes1-B44 rec8-2A<<Kan rec12::hyg natMX6-3pk-moa1-S143K T150K
PM12	h⁺ mes1-B44 rec8-2A<<Kan rec12::hyg natMX6-3pk-moa1-S143K T150K
PM21	h^- leu1 imr1L-GFP mes1-B44 rec8-2A<<Kan rec12::hyg natMX6-3pk-moa1-S143D T150E
PM22	h⁺ mes1-B44 rec8-2A<<Kan rec12::hyg natMX6-3pk-moa1-S143D T150E
PM31	h^- leu1 imr1L-GFP mes1-B44 rec8-2A<<Kan rec12::hyg natMX6-3pk-moa1-E165K
PM32	h⁺ mes1-B44 rec8-2A<<Kan rec12::hyg natMX6-3pk-moa1-E165K
PM41	h^- leu1 imr1L-GFP mes1-B44 rec8-2A<<Kan rec12::hyg natMX6-3pk-moa1-D167K E168K
PM42	h⁺ mes1-B44 rec8-2A<<Kan rec12::hyg natMX6-3pk-moa1-D167K E168K
PM51	h^- leu1 imr1L-GFP mes1-B44 rec8-2A<<Kan rec12::hyg natMX6-3pk-moa1-I170K L171K
PM52	h⁺ mes1-B44 rec8-2A<<Kan rec12::hyg natMX6-3pk-moa1-I170K L171K
LM1	h^- leu1imr1L-GFP mes1-B44 rec8-2A<<Kan rec12::bsdr moa1::natr
LM2	h⁺ mes1-B44 rec8-2A<<Kan rec12::bsdr moa1::natr
PM-W	h⁹⁰ mei4::hyg natMX6-GFP-3pk-moa1
PM36	h⁹⁰ mei4::hyg natMX6-GFP-3pk-moa1-E165K
PM37	h⁹⁰ mei4::hyg natMX6-GFP-3pk-moa1-D167K E168K
PM38	h⁹⁰ mei4::hyg natMX6-GFP-3pk-moa1-I170K L171K

Moa1与CENP-C和Rec8的相互作用及其在裂殖酵母减数分裂中的功能

Functional roles of the interaction of Moa1 with CENP-C and Rec8 in meiosis of Schizosaccharomyces pombe

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