Functional analysis of Cdc20 reveals a critical role of CRY box in mitotic checkpoint signaling
Functional analysis of Cdc20 reveals a critical role of CRY box in mitotic checkpoint signaling
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DOI：
10.1101/2023.07.26.550666
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Gang Zhang
中科院分区：
文献类型：
--
作者：
Yuqing Zhang;Rose Young;Dimitriya H Garvanska;Chunlin Song;Yujing Zhai;Ying Wang;Hongfei Jiang;Jing Fang;Jakob Nilsson;Claudio Alfieri;Gang Zhang
Accurate chromosome segregation is coordinated by the spindle assembly checkpoint.(SAC) through its effector the mitotic checkpoint complex (MCC), to inhibit the.anaphase-promoting complex or cyclosome (APC/C). Cdc20 is an essential mitotic.regulator since it promotes mitotic exit through activating the APC/C and monitors.kinetochore-microtubule attachment through activating the SAC. The proper.functioning of Cdc20 requires multiple interactions with APC/C and MCC subunits. To.functionally assess each of these interactions within cells requires efficient depletion of.endogenous Cdc20, which is highly difficult to achieve by RNAi. Here we generated.Cdc20 RNAi sensitive cell lines by CRISPR/Cas9 which display a penetrant metaphase.arrest phenotype 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 to be critical for.the SAC by promoting the MCC formation and stabilizing the interaction between the.MCC and APC/C. These data reveal additional regulatory components within the SAC.and establish a novel method to interrogate Cdc20 function.
