title: And-1 coordinates with Claspin for efficient Chk1 activation in response to replication stress.
abstract: The replisome is important for DNA replication checkpoint activation, but how specific components of the replisome coordinate with ATR to activate Chk1 in human cells remains largely unknown. Here, we demonstrate that And-1, a replisome component, acts together with ATR to activate Chk1. And-1 is phosphorylated at T826 by ATR following replication stress, and this phosphorylation is required for And-1 to accumulate at the damage sites, where And-1 promotes the interaction between Claspin and Chk1, thereby stimulating efficient Chk1 activation by ATR. Significantly, And-1 binds directly to ssDNA and facilitates the association of Claspin with ssDNA. Furthermore, And-1 associates with replication forks and is required for the recovery of stalled forks. These studies establish a novel ATR-And-1 axis as an important regulator for efficient Chk1 activation and reveal a novel mechanism of how the replisome regulates the replication checkpoint and genomic stability.
authors.affiliation: Department of Biochemistry and Molecular Medicine, The George Washington University Medical School, Washington, DC, USA.
authors.forename: Jing
authors.initials: J
authors.lastname: Hao
authors.affiliation: National Institute of Child Health and Human Development, Bethesda, MD, USA.
authors.forename: Christelle
authors.initials: C
authors.lastname: de Renty
authors.affiliation: Department of Biochemistry and Molecular Medicine, The George Washington University Medical School, Washington, DC, USA.
authors.forename: Yongming
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Biochemistry and Molecular Medicine, The George Washington University Medical School, Washington, DC, USA.
authors.forename: Haijie
authors.initials: H
authors.lastname: Xiao
authors.affiliation: Department of Biological Sciences, Florida Institute of Technology, Melbourne, FL, USA.
authors.forename: Michael G
authors.initials: MG
authors.lastname: Kemp
authors.affiliation: Department of Biochemistry and Molecular Medicine, The George Washington University Medical School, Washington, DC, USA.
authors.forename: Zhiyong
authors.initials: Z
authors.lastname: Han
authors.affiliation: National Institute of Child Health and Human Development, Bethesda, MD, USA.
authors.forename: Melvin L
authors.initials: ML
authors.lastname: DePamphilis
authors.affiliation: Department of Biochemistry and Molecular Medicine, The George Washington University Medical School, Washington, DC, USA wz6812@gwu.edu.
authors.forename: Wenge
authors.initials: W
authors.lastname: Zhu
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D000906:Antibodies / Q000276:immunology; D000071877:Checkpoint Kinase 1; D004261:DNA Replication / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D057809:HEK293 Cells; D006801:Humans; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D008954:Models, Biological; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000906:Antibodies; C488158:CLSPN protein, human; D004268:DNA-Binding Proteins; D034741:RNA, Small Interfering; C544202:WDHD1 protein, human; D011494:Protein Kinases; C000605867:CHEK1 protein, human; D000071877:Checkpoint Kinase 1
keywords: ATR; And‐1; Chk1; Claspin; replication stress