Mutation of aspartic acid 199 in USP1 disrupts its deubiquitinating activity and impairs DNA repair

Abstract

The deubiquitinating enzyme USP1 contains highly conserved motifs forming its catalytic center. Recently, the COSMIC mutation database identified a mutation in USP1 at Asp‐199 in endometrial cancer. Here, we investigated the role of Asp‐199 for USP1 function. The mutation of aspartic acid to alanine (D199A) resulted in failure of USP1 to undergo autocleavage and form a complex with ubiquitin, indicating D199A Usp1 is catalytically inactive. The D199A mutation did not affect the interaction with Uaf1. Moreover, D199A Usp1 had defects in deubiquitination of FANCD2 and PCNA and displayed reduced FANCD2 foci formation and DNA repair efficiency. Furthermore, mutation of Asp‐199 to glutamic acid resulted in phenotypes similar to the D199A mutation. Collectively, our findings demonstrate the importance of Asp‐199 for USP1 activity and suggest the implications of USP1 downregulation in cancer.

Association of environmental exposure to chromium with differential DNA methylation: An epigenome-wide study

“…The most significant Cr-related CpG, cg20690919, is located in the gene ubiquitin specific peptidase 1 ( USP1 ) gene, which is a 785 amino-acid deubiquitinating enzyme with His and Cys domains ( Jang and Kim, 2021 ). USP1 can involve in DNA interstrand crosslink repair by deubiquitinating the Fanconi anemia protein ( Taniguchi et al, 2002 ; Kim and D'Andrea, 2012 ) and regulating DNA damage response pathways ( Jang and Kim, 2021 ).…”

“…The most significant Cr-related CpG, cg20690919, is located in the gene ubiquitin specific peptidase 1 ( USP1 ) gene, which is a 785 amino-acid deubiquitinating enzyme with His and Cys domains ( Jang and Kim, 2021 ). USP1 can involve in DNA interstrand crosslink repair by deubiquitinating the Fanconi anemia protein ( Taniguchi et al, 2002 ; Kim and D'Andrea, 2012 ) and regulating DNA damage response pathways ( Jang and Kim, 2021 ). USP1-associated factor 1 ( UAF1 ) and RAD51 -associated protein 1 ( RAD51AP1 ) enhance the activity of recombinase RAD51 during DNA repair through the homologous recombination pathway ( Liang et al, 2020 ).…”