FANCD2-dependent mitotic DNA synthesis relies on PCNA K164 ubiquitination
Wendy Leung,
Ryan M. Baxley,
Emma Traband,
Ya-Chu Chang,
Colette B. Rogers,
Liangjun Wang,
Wesley Durrett,
Kendall S. Bromley,
Lidia Fiedorowicz,
Tanay Thakar,
Anika Tella,
Alexandra Sobeck,
Eric A. Hendrickson,
George-Lucian Moldovan,
Naoko Shima,
Anja-Katrin Bielinsky
Affiliations
Wendy Leung
Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA
Ryan M. Baxley
Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA
Emma Traband
Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN 55455, USA
Ya-Chu Chang
Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA
Colette B. Rogers
Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA
Liangjun Wang
Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA
Wesley Durrett
Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA
Kendall S. Bromley
Department of Biochemistry and Molecular Genetics, University of Virginia, Charlottesville, VA 22903, USA
Lidia Fiedorowicz
Department of Biochemistry and Molecular Genetics, University of Virginia, Charlottesville, VA 22903, USA
Tanay Thakar
Department of Biochemistry and Molecular Biology, The Pennsylvania State University College of Medicine, Hershey, PA 17033, USA
Anika Tella
Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN 55455, USA
Alexandra Sobeck
Institute for Human Genetics, Biocenter, University of Würzburg, Würzburg, Germany
Eric A. Hendrickson
Department of Medicine, University of Virginia, Charlottesville, VA 22903, USA
George-Lucian Moldovan
Department of Biochemistry and Molecular Biology, The Pennsylvania State University College of Medicine, Hershey, PA 17033, USA
Naoko Shima
Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN 55455, USA; Corresponding author
Anja-Katrin Bielinsky
Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA; Department of Biochemistry and Molecular Genetics, University of Virginia, Charlottesville, VA 22903, USA; Corresponding author
Summary: Ubiquitination of proliferating cell nuclear antigen (PCNA) at lysine 164 (K164) activates DNA damage tolerance pathways. Currently, we lack a comprehensive understanding of how PCNA K164 ubiquitination promotes genome stability. To evaluate this, we generated stable cell lines expressing PCNAK164R from the endogenous PCNA locus. Our data reveal that the inability to ubiquitinate K164 causes perturbations in global DNA replication. Persistent replication stress generates under-replicated regions and is exacerbated by the DNA polymerase inhibitor aphidicolin. We show that these phenotypes are due, in part, to impaired Fanconi anemia group D2 protein (FANCD2)-dependent mitotic DNA synthesis (MiDAS) in PCNAK164R cells. FANCD2 mono-ubiquitination is significantly reduced in PCNAK164R mutants, leading to reduced chromatin association and foci formation, both prerequisites for FANCD2-dependent MiDAS. Furthermore, K164 ubiquitination coordinates direct PCNA/FANCD2 colocalization in mitotic nuclei. Here, we show that PCNA K164 ubiquitination maintains human genome stability by promoting FANCD2-dependent MiDAS to prevent the accumulation of under-replicated DNA.
