title: HUWE1 interacts with PCNA to alleviate replication stress.
abstract: Defects in DNA replication, DNA damage response, and DNA repair compromise genomic stability and promote cancer development. In particular, unrepaired DNA lesions can arrest the progression of the DNA replication machinery during S-phase, causing replication stress, mutations, and DNA breaks. HUWE1 is a HECT-type ubiquitin ligase that targets proteins involved in cell fate, survival, and differentiation. Here, we report that HUWE1 is essential for genomic stability, by promoting replication of damaged DNA We show that HUWE1-knockout cells are unable to mitigate replication stress, resulting in replication defects and DNA breakage. Importantly, we find that this novel role of HUWE1 requires its interaction with the replication factor PCNA, a master regulator of replication fork restart, at stalled replication forks. Finally, we provide evidence that HUWE1 mono-ubiquitinates H2AX to promote signaling at stalled forks. Altogether, our work identifies HUWE1 as a novel regulator of the replication stress response.
authors.affiliation: Department of Biochemistry and Molecular Biology, The Pennsylvania State University College of Medicine, Hershey, PA, USA.
authors.forename: Katherine N
authors.initials: KN
authors.lastname: Choe
authors.affiliation: Department of Biochemistry and Molecular Biology, The Pennsylvania State University College of Medicine, Hershey, PA, USA.
authors.forename: Claudia M
authors.initials: CM
authors.lastname: Nicolae
authors.affiliation: Department of Biochemistry and Molecular Biology, The Pennsylvania State University College of Medicine, Hershey, PA, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Constantin
authors.affiliation: Department of Biochemistry and Molecular Biology, The Pennsylvania State University College of Medicine, Hershey, PA, USA Department of Pharmacology, The Pennsylvania State University College of Medicine, Hershey, PA, USA Institute for Personalized Medicine, The Pennsylvania State University College of Medicine, Hershey, PA, USA.
authors.forename: Yuka
authors.initials: Y
authors.lastname: Imamura Kawasawa
authors.affiliation: Unidad de Investigación, Hospital Universitario de Canarias, Instituto de Tecnologías Biomédicas, La Laguna Tenerife, Spain.
authors.forename: Maria Rocio
authors.initials: MR
authors.lastname: Delgado-Diaz
authors.affiliation: Department of Medicine, University of Colorado School of Medicine, Aurora, CO, USA Department of Biostatistics and Informatics, Colorado School of Public Health, Aurora, CO, USA Molecular Oncology Program, University of Colorado Cancer Center, Aurora, CO, USA.
authors.forename: Subhajyoti
authors.initials: S
authors.lastname: De
authors.affiliation: Unidad de Investigación, Hospital Universitario de Canarias, Instituto de Tecnologías Biomédicas, La Laguna Tenerife, Spain.
authors.forename: Raimundo
authors.initials: R
authors.lastname: Freire
authors.affiliation: Unidad de Investigación, Hospital Universitario de Canarias, Instituto de Tecnologías Biomédicas, La Laguna Tenerife, Spain.
authors.forename: Veronique Aj
authors.initials: VA
authors.lastname: Smits
authors.affiliation: Department of Biochemistry and Molecular Biology, The Pennsylvania State University College of Medicine, Hershey, PA, USA gmoldovan@hmc.psu.edu.
authors.forename: George-Lucian
authors.identifier: 0000-0003-3825-149X
authors.initials: GL
authors.lastname: Moldovan
mesh_terms: D002460:Cell Line; D004249:DNA Damage; D004260:DNA Repair; D004261:DNA Replication; D055786:Gene Knockout Techniques; D042822:Genomic Instability; D006657:Histones / Q000378:metabolism; D006801:Humans; D010641:Phenotype; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism*; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D013312:Stress, Physiological; D025521:Tumor Suppressor Proteins; D025801:Ubiquitin / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism*; D054875:Ubiquitination
chemical_list: D006657:Histones; D018809:Proliferating Cell Nuclear Antigen; D025521:Tumor Suppressor Proteins; D025801:Ubiquitin; C502830:HUWE1 protein, human; D044767:Ubiquitin-Protein Ligases
keywords: DNA replication; H2AX; HUWE1; PCNA; genomic instability