title: The structure and polymerase-recognition mechanism of the crucial adaptor protein AND-1 in the human replisome.
abstract: DNA replication in eukaryotic cells is performed by a multiprotein complex called the replisome, which consists of helicases, polymerases, and adaptor molecules. HumanacidicnucleoplasmicDNA-binding protein 1 (AND-1), also known as WD repeat and high mobility group (HMG)-box DNA-binding protein 1 (WDHD1), is an adaptor molecule crucial for DNA replication. Although structural information for the AND-1 yeast ortholog is available, the mechanistic details for how human AND-1 protein anchors the lagging-strand DNA polymerase α (pol α) to the DNA helicase complex (Cdc45-MCM2-7-GINS, CMG) await elucidation. Here, we report the structures of the N-terminal WD40 and SepB domains of human AND-1, as well as a biochemical analysis of the C-terminal HMG domain. We show that AND-1 exists as a homotrimer mediated by the SepB domain. Mutant study results suggested that a positively charged groove within the SepB domain provides binding sites for pol α. Different from its ortholog protein in budding yeast, human AND-1 is recruited to the CMG complex, mediated by unknown participants other than Go Ichi Ni San. In addition, we show that AND-1 binds to DNAin vitro, using its C-terminal HMG domain. In conclusion, our findings provide important insights into the mechanistic details of human AND-1 function, advancing our understanding of replisome formation during eukaryotic replication.
authors.affiliation: From the National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101 and.|the College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Chengcheng
authors.initials: C
authors.lastname: Guan
authors.affiliation: From the National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101 and.|the College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.affiliation: From the National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101 and.
authors.forename: Dapeng
authors.initials: D
authors.lastname: Sun
authors.affiliation: From the National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101 and liuyingfang@ibp.ac.cn.
authors.forename: Yingfang
authors.initials: Y
authors.lastname: Liu
authors.affiliation: From the National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101 and hhliang@moon.ibp.ac.cn.
authors.forename: Huanhuan
authors.initials: H
authors.lastname: Liang
mesh_terms: D004247:DNA / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D004265:DNA Helicases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004256:DNA Polymerase I / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004261:DNA Replication / Q000502:physiology; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D009097:Multienzyme Complexes / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000072417:Protein Domains; D055503:Protein Multimerization
chemical_list: D004268:DNA-Binding Proteins; D009097:Multienzyme Complexes; C544202:WDHD1 protein, human; D004247:DNA; D004256:DNA Polymerase I; D004265:DNA Helicases
keywords: AND-1; DNA polymerase; DNA replication; DNA-protein interaction; HMG domain; crystal structure; polymerase a; protein-protein interaction; replisome