Functional analysis of Cdc20 reveals a critical role of CRY box in mitotic checkpoint signaling

Yuqing Zhang; Rose Young; Dimitriya H. Garvanska; Chunlin Song; Yujing Zhai; Ying Wang; Hongfei Jiang; Jing Fang; Jakob Nilsson; Claudio Alfieri; Gang Zhang

doi:10.1038/s42003-024-05859-6

Functional analysis of Cdc20 reveals a critical role of CRY box in mitotic checkpoint signaling

Yuqing Zhang, Rose Young, Dimitriya H. Garvanska, Chunlin Song, Yujing Zhai, Ying Wang, Hongfei Jiang, Jing Fang, Jakob Nilsson, Claudio Alfieri, Gang Zhang^*

^*Corresponding author af dette arbejde

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Abstract

Accurate mitosis is coordinated by the spindle assembly checkpoint (SAC) through the mitotic checkpoint complex (MCC), which inhibits the anaphase-promoting complex or cyclosome (APC/C). As an essential regulator, Cdc20 promotes mitotic exit through activating APC/C and monitors kinetochore-microtubule attachment through activating SAC. Cdc20 requires multiple interactions with APC/C and MCC subunits to elicit these functions. Functionally assessing these interactions within cells requires efficient depletion of endogenous Cdc20, which is highly difficult to achieve by RNA interference (RNAi). Here we generated Cdc20 RNAi-sensitive cell lines which display a penetrant metaphase arrest by a single RNAi treatment. In this null background, we accurately measured the contribution of each known motif of Cdc20 on APC/C and SAC activation. The CRY box, a previously identified degron, was found critical for SAC by promoting MCC formation and its interaction with APC/C. These data reveal additional regulation within the SAC and establish a novel method to interrogate Cdc20.

Originalsprog	Engelsk
Artikelnummer	164
Tidsskrift	Communications Biology
Vol/bind	7
Antal sider	12
ISSN	2399-3642
DOI	https://doi.org/10.1038/s42003-024-05859-6
Status	Udgivet - 2024

Bibliografisk note

Funding Information:
G.Z. is supported by the National Natural Science Foundation of China (31970666) and the Taishan Scholar Project (tsqn201812054) from Shandong, China. C.A. and R.Y. are supported by the Sir Henry Dale Fellowship 215458/Z/19/Z. R.Y. is also supported by the Institute of Cancer Research (ICR), and the grant number allocated is GFR005X. We acknowledge Jing Yang, Ziguo Zhang, and David Barford for helping with the APC/C baculovirus generation.

Publisher Copyright:
© The Author(s) 2024.

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Citationsformater

Functional analysis of Cdc20 reveals a critical role of CRY box in mitotic checkpoint signaling. / Zhang, Yuqing; Young, Rose; Garvanska, Dimitriya H.; Song, Chunlin; Zhai, Yujing; Wang, Ying; Jiang, Hongfei; Fang, Jing; Nilsson, Jakob; Alfieri, Claudio; Zhang, Gang.

I: Communications Biology , Bind 7, 164, 2024.

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

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abstract = "Accurate mitosis is coordinated by the spindle assembly checkpoint (SAC) through the mitotic checkpoint complex (MCC), which inhibits the anaphase-promoting complex or cyclosome (APC/C). As an essential regulator, Cdc20 promotes mitotic exit through activating APC/C and monitors kinetochore-microtubule attachment through activating SAC. Cdc20 requires multiple interactions with APC/C and MCC subunits to elicit these functions. Functionally assessing these interactions within cells requires efficient depletion of endogenous Cdc20, which is highly difficult to achieve by RNA interference (RNAi). Here we generated Cdc20 RNAi-sensitive cell lines which display a penetrant metaphase arrest by a single RNAi treatment. In this null background, we accurately measured the contribution of each known motif of Cdc20 on APC/C and SAC activation. The CRY box, a previously identified degron, was found critical for SAC by promoting MCC formation and its interaction with APC/C. These data reveal additional regulation within the SAC and establish a novel method to interrogate Cdc20.",

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T1 - Functional analysis of Cdc20 reveals a critical role of CRY box in mitotic checkpoint signaling

AU - Zhang, Yuqing

AU - Young, Rose

AU - Garvanska, Dimitriya H.

AU - Song, Chunlin

AU - Zhai, Yujing

AU - Wang, Ying

AU - Jiang, Hongfei

AU - Fang, Jing

AU - Nilsson, Jakob

AU - Alfieri, Claudio

AU - Zhang, Gang

PY - 2024

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N2 - Accurate mitosis is coordinated by the spindle assembly checkpoint (SAC) through the mitotic checkpoint complex (MCC), which inhibits the anaphase-promoting complex or cyclosome (APC/C). As an essential regulator, Cdc20 promotes mitotic exit through activating APC/C and monitors kinetochore-microtubule attachment through activating SAC. Cdc20 requires multiple interactions with APC/C and MCC subunits to elicit these functions. Functionally assessing these interactions within cells requires efficient depletion of endogenous Cdc20, which is highly difficult to achieve by RNA interference (RNAi). Here we generated Cdc20 RNAi-sensitive cell lines which display a penetrant metaphase arrest by a single RNAi treatment. In this null background, we accurately measured the contribution of each known motif of Cdc20 on APC/C and SAC activation. The CRY box, a previously identified degron, was found critical for SAC by promoting MCC formation and its interaction with APC/C. These data reveal additional regulation within the SAC and establish a novel method to interrogate Cdc20.

AB - Accurate mitosis is coordinated by the spindle assembly checkpoint (SAC) through the mitotic checkpoint complex (MCC), which inhibits the anaphase-promoting complex or cyclosome (APC/C). As an essential regulator, Cdc20 promotes mitotic exit through activating APC/C and monitors kinetochore-microtubule attachment through activating SAC. Cdc20 requires multiple interactions with APC/C and MCC subunits to elicit these functions. Functionally assessing these interactions within cells requires efficient depletion of endogenous Cdc20, which is highly difficult to achieve by RNA interference (RNAi). Here we generated Cdc20 RNAi-sensitive cell lines which display a penetrant metaphase arrest by a single RNAi treatment. In this null background, we accurately measured the contribution of each known motif of Cdc20 on APC/C and SAC activation. The CRY box, a previously identified degron, was found critical for SAC by promoting MCC formation and its interaction with APC/C. These data reveal additional regulation within the SAC and establish a novel method to interrogate Cdc20.

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