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bioRxiv · 10.1101/2022.11.10.515958

Transcription-independent hold of the G1/S transition is exploited to cope with DNA replication stress

Jin, Y.·lan, G.·Zhang, J.·Sun, H.·Li, X.·Cao, Q.·Tang, C.·Yang, X.·Lou, H.·Hou, W.
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Abstract

RB1 (retinoblastoma) members control the G1/S commitment as transcriptional repressors in eukaryotic cells. Here we uncover that an extra copy of RB1 equivalent (WHI7 or WHI5) is sufficient to bypass the indispensability of the central genomic checkpoint kinases Mec1ATR-Rad53CHK1 in Saccharomyces cerevisiae. Mec1-Rad53 directly phosphorylate Whi7/5, antagonizing their nuclear export or protein turnover upon replication stress. Through in vitro reconstitution, we show that Whi7 C-terminus directly binds and hinders S-CDK-Cks1 from processively phosphorylating Sic1. By microfluidic single-cell real-time quantitative imaging, we demonstrate that both Whi7 and Whi5 are required to flatten the degradation curve of the major S-CDK inhibitor Sic1 in vivo. These findings reveal an eclipsed transcription-independent role of Whi7 homologs, which is highlighted by genome integrity checkpoints to hold the G1/S transition instantly as a rapid response to unforeseeable replication threats. Key pointsO_LIWhi7 overexpression bypasses the essential function of Mec1 and Rad53 in a transcription-independent way. C_LIO_LIWhi7 is stabilized by checkpoint-mediated phosphorylation. C_LIO_LIWhi7 binds and hinders S-CDK-Cks1 from multi-phosphorylation of Sci1, thereby prolonging Sic1 degradation and G1/S transition. C_LI

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Jin, Y., lan, G., Zhang, J., Sun, H., Li, X., Cao, Q., Tang, C., Yang, X., Lou, H., Hou, W.. 2022-11-11. Transcription-independent hold of the G1/S transition is exploited to cope with DNA replication stress. https://doi.org/10.1101/2022.11.10.515958

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