title: HERC2 Interacts with Claspin and regulates DNA origin firing and replication fork progression.
abstract: DNA replication, recombination, and repair are highly interconnected processes the disruption of which must be coordinated in cancer. HERC2, a large HECT protein required for homologous recombination repair, is an E3 ubiquitin ligase that targets breast cancer suppressor BRCA1 for degradation. Here, we show that HERC2 is a component of the DNA replication fork complex that plays a critical role in DNA elongation and origin firing. In the presence of BRCA1, endogenous HERC2 interacts with Claspin, a protein essential for G(2)-M checkpoint activation and replication fork stability. Claspin depletion slowed S-phase progression and additional HERC2 depletion reduced the effect of Claspin depletion. In addition, HERC2 interacts with replication fork complex proteins. Depletion of HERC2 alleviated the slow replication fork progression in Claspin-deficient cells, suppressed enhanced origin firing, and led to a decrease in MCM2 phosphorylation. In a HERC2-dependent manner, treatment of cells with replication inhibitor aphidicolin enhanced MCM2 phosphorylation. Taken together, our results suggest that HERC2 regulates DNA replication progression and origin firing by facilitating MCM2 phosphorylation. These findings establish HERC2 as a critical function in DNA repair, checkpoint activation, and DNA replication.
authors.affiliation: Department of Translational Oncology, Gastroenterology and Hepatology, Clinical Oncology, and Breast and Endocrine Surgery, Institute of Advanced Medical Science, St. Marianna University Graduate School of Medicine, Kawasaki, Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Izawa
authors.forename: Wenwen
authors.initials: W
authors.lastname: Wu
authors.forename: Ko
authors.initials: K
authors.lastname: Sato
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Nishikawa
authors.forename: Akihiro
authors.initials: A
authors.lastname: Kato
authors.forename: Narikazu
authors.initials: N
authors.lastname: Boku
authors.forename: Fumio
authors.initials: F
authors.lastname: Itoh
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Ohta
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D019313:BRCA1 Protein / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D004260:DNA Repair; D004261:DNA Replication; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism*; D006801:Humans; D064111:Minichromosome Maintenance Complex Component 2; D009687:Nuclear Proteins / Q000378:metabolism; D010766:Phosphorylation; D018741:Replication Origin; D016196:S Phase; D044767:Ubiquitin-Protein Ligases
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D019313:BRCA1 Protein; C488158:CLSPN protein, human; D018797:Cell Cycle Proteins; D020662:Guanine Nucleotide Exchange Factors; D009687:Nuclear Proteins; C118566:HERC2 protein, human; D044767:Ubiquitin-Protein Ligases; C084859:MCM2 protein, human; D064111:Minichromosome Maintenance Complex Component 2