Protein kinase ATR inhibits E3 ubiquitin ligase CRL4PRL1 to stabilize ribonucleotide reductase in response to replication stress

Weiyi Bao; Weijia Zhang; Yongchi Huang; Yan Zhao; Cong Wu; Leilei Duan; Lili Wang; Shunping Yan

doi:10.1016/j.celrep.2023.112685

Protein kinase ATR inhibits E3 ubiquitin ligase CRL4^PRL1 to stabilize ribonucleotide reductase in response to replication stress

Cell Rep. 2023 Jul 25;42(7):112685. doi: 10.1016/j.celrep.2023.112685. Epub 2023 Jun 23.

Authors

Weiyi Bao¹, Weijia Zhang¹, Yongchi Huang¹, Yan Zhao¹, Cong Wu¹, Leilei Duan¹, Lili Wang², Shunping Yan³

Affiliations

¹ Hubei Hongshan Laboratory, Wuhan, Hubei 430070, China; College of Life Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, China; Shenzhen Institute of Nutrition and Health, Huazhong Agricultural University, Shenzhen 518000, China; Shenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Shenzhen 518000, China; Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518000, China.
² Hubei Hongshan Laboratory, Wuhan, Hubei 430070, China; College of Life Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, China; Shenzhen Institute of Nutrition and Health, Huazhong Agricultural University, Shenzhen 518000, China; Shenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Shenzhen 518000, China; Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518000, China. Electronic address: liliwang@mail.hzau.edu.cn.
³ Hubei Hongshan Laboratory, Wuhan, Hubei 430070, China; College of Life Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, China; Shenzhen Institute of Nutrition and Health, Huazhong Agricultural University, Shenzhen 518000, China; Shenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Shenzhen 518000, China; Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518000, China. Electronic address: spyan@mail.hzau.edu.cn.

PMID: 37354461
DOI: 10.1016/j.celrep.2023.112685

Abstract

The protein kinase ATR is essential for replication stress responses in all eukaryotes. Ribonucleotide reductase (RNR) catalyzes the formation of deoxyribonucleotide (dNTP), the universal building block for DNA replication and repair. However, the relationship between ATR and RNR is not well understood. Here, we show that ATR promotes the protein stability of RNR in Arabidopsis. Through an activation tagging-based genetic screen, we found that overexpression of TSO2, a small subunit of RNR, partially suppresses the hypersensitivity of the atr mutant to replication stress. Biochemically, TSO2 interacts with PRL1, a central subunit of the Cullin4-based E3 ubiquitin ligase CRL4^PRL1, which polyubiquitinates TSO2 and promotes its degradation. ATR inhibits CRL4^PRL1 to attenuate TSO2 degradation. Our work provides an important insight into the replication stress responses and a post-translational regulatory mechanism for RNR. Given the evolutionary conservation of the proteins involved, the ATR-PRL1-RNR module may act across eukaryotes.

Keywords: ATR; CP: Molecular biology; DNA damage response; DNA replication stress; PRL1; TSO2; WEE1; ribonucleotide reductase.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Arabidopsis Proteins* / genetics
Arabidopsis Proteins* / metabolism
Arabidopsis* / genetics
Arabidopsis* / metabolism
Ataxia Telangiectasia Mutated Proteins / metabolism
DNA Damage
DNA Replication
Ribonucleotide Reductases* / genetics
Ribonucleotide Reductases* / metabolism
Ubiquitin-Protein Ligases / genetics
Ubiquitin-Protein Ligases / metabolism

Substances

Arabidopsis Proteins
Ataxia Telangiectasia Mutated Proteins
ATR protein, Arabidopsis
Ribonucleotide Reductases
Ubiquitin-Protein Ligases
PRL1 protein, plant