Reconstitution of an active human CENP-E motor.
Reconstitution of an active human CENP-E motor.
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DOI：
10.1098/rsob.210389
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发表时间：
2022-03
期刊：
影响因子：
5.8
通讯作者：
Welburn JPI
中科院分区：
文献类型：
--
作者：
Craske B;Legal T;Welburn JPI
CENP-E是一种大型运动蛋白，通过促进染色体捕获和排列，促进纺锤体中的微管通量，在有丝分裂中起关键作用。到目前为止，还不可能获得活性的人CENP-E来研究其分子性质。对爪蟾的CENP-E马达进行了体外表征，并作为模型马达；然而，它的蛋白质序列与人类的CENP-E有很大的不同。在这里，我们在体外表征了人类CENP-E的运动性。与全长CENP-E马达截断相比，全长CENP-E在微管上的运行长度增加，停留时间更长，这表明当全长马达激活时，c端微管结合位点增强了加工能力。与构成活性的人CENP-E截断相比，全长人CENP-E在体外的微管着陆率降低，这表明非运动性卷曲线圈区域自我调节运动活动。总之，我们证明了人类CENP-E是一个进程马达，为研究人类CENP-E在人类细胞分裂过程中如何驱动染色体聚集和纺锤体组织的机制基础提供了有用的工具。
CENP-E is a large kinesin motor protein which plays pivotal roles in mitosis by facilitating chromosome capture and alignment, and promoting microtubule flux in the spindle. So far, it has not been possible to obtain active human CENP-E to study its molecular properties. Xenopus CENP-E motor has been characterized in vitro and is used as a model motor; however, its protein sequence differs significantly from human CENP-E. Here, we characterize human CENP-E motility in vitro. Full-length CENP-E exhibits an increase in run length and longer residency times on microtubules when compared to CENP-E motor truncations, indicating that the C-terminal microtubule-binding site enhances the processivity when the full-length motor is active. In contrast with constitutively active human CENP-E truncations, full-length human CENP-E has a reduced microtubule landing rate in vitro, suggesting that the non-motor coiled-coil regions self-regulate motor activity. Together, we demonstrate that human CENP-E is a processive motor, providing a useful tool to study the mechanistic basis for how human CENP-E drives chromosome congression and spindle organization during human cell division.
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影响因子：
14.8
作者：
Gietz, R. Daniel;Schiestl, Robert H.
通讯作者：
Schiestl, Robert H.
影响因子：
64.8
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Jumper J;Evans R;Pritzel A;Green T;Figurnov M;Ronneberger O;Tunyasuvunakool K;Bates R;Žídek A;Potapenko A;Bridgland A;Meyer C;Kohl SAA;Ballard AJ;Cowie A;Romera-Paredes B;Nikolov S;Jain R;Adler J;Back T;Petersen S;Reiman D;Clancy E;Zielinski M;Steinegger M;Pacholska M;Berghammer T;Bodenstein S;Silver D;Vinyals O;Senior AW;Kavukcuoglu K;Kohli P;Hassabis D
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Hassabis D
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21.3
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Coy, DL;Hancock, WO;Howard, J
通讯作者：
Howard, J
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64.5
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Case, RB;Pierce, DW;Vale, RD
通讯作者：
Vale, RD
影响因子：
4
作者：
Legal, Thibault;Hayward, Daniel;Welburn, Julie P. I.
通讯作者：
Welburn, Julie P. I.