title: A single aspartate mutation in the conserved catalytic site of Rev3L generates a hypomorphic phenotype in vivo and in vitro.
abstract: Rev3, the catalytic subunit of yeast DNA polymerase ζ, is required for UV resistance and UV-induced mutagenesis, while its mammalian ortholog, REV3L, plays further vital roles in cell proliferation and embryonic development. To assess the contribution of REV3L catalytic activity to its in vivo function, we generated mutant mouse strains in which one or two Ala residues were substituted to the Asp of the invariant catalytic YGDTDS motif. The simultaneous mutation of both Asp (ATA) phenocopies the Rev3l knockout, which proves that the catalytic activity is mandatory for the vital functions of Rev3L, as reported recently. Surprisingly, although the mutation of the first Asp severely impairs the enzymatic activity of other B-family DNA polymerases, the corresponding mutation of Rev3 (ATD) is hypomorphic in yeast and mouse, as it does not affect viability and proliferation and moderately impacts UVC-induced cell death and mutagenesis. Interestingly, Rev3l hypomorphic mutant mice display a distinct, albeit modest, alteration of the immunoglobulin gene mutation spectrum at G-C base pairs, further documenting its role in this process.
authors.affiliation: Institut Necker-Enfants Malades, INSERM U1151-CNRS UMR 8253, Faculté de Médecine Paris Descartes, Université Paris Descartes, Sorbonne Paris Cité, Paris, France. Electronic address: r.fritzen@dundee.ac.uk.
authors.forename: Rémi
authors.initials: R
authors.lastname: Fritzen
authors.affiliation: Institut Necker-Enfants Malades, INSERM U1151-CNRS UMR 8253, Faculté de Médecine Paris Descartes, Université Paris Descartes, Sorbonne Paris Cité, Paris, France. Electronic address: Frederic.Delbos@univ-nantes.fr.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Delbos
authors.affiliation: Institut Necker-Enfants Malades, INSERM U1151-CNRS UMR 8253, Faculté de Médecine Paris Descartes, Université Paris Descartes, Sorbonne Paris Cité, Paris, France. Electronic address: annie.de-smet@inserm.fr.
authors.forename: Annie
authors.initials: A
authors.lastname: De Smet
authors.affiliation: Institut Jacques Monod, CNRS UMR 7592, Université Paris Diderot, Sorbonne Paris Cité, Paris, France. Electronic address: Benoit.PALANCADE@ijm.fr.
authors.forename: Benoît
authors.initials: B
authors.lastname: Palancade
authors.affiliation: Department of Pharmacology, University of Michigan, Ann Arbor, MI, USA. Electronic address: ccanman@umich.edu.
authors.forename: Christine E
authors.initials: CE
authors.lastname: Canman
authors.affiliation: Institut Gustave Roussy, CNRS UMR 8200, Villejuif, and Université Paris-Sud, Orsay, France. Electronic address: Said.AOUFOUCHI@gustaveroussy.fr.
authors.forename: Said
authors.initials: S
authors.lastname: Aoufouchi
authors.affiliation: Institut Necker-Enfants Malades, INSERM U1151-CNRS UMR 8253, Faculté de Médecine Paris Descartes, Université Paris Descartes, Sorbonne Paris Cité, Paris, France. Electronic address: jean-claude.weill@inserm.fr.
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Weill
authors.affiliation: Institut Necker-Enfants Malades, INSERM U1151-CNRS UMR 8253, Faculté de Médecine Paris Descartes, Université Paris Descartes, Sorbonne Paris Cité, Paris, France. Electronic address: claude-agnes.reynaud@inserm.fr.
authors.forename: Claude-Agnès
authors.initials: CA
authors.lastname: Reynaud
authors.affiliation: Institut Necker-Enfants Malades, INSERM U1151-CNRS UMR 8253, Faculté de Médecine Paris Descartes, Université Paris Descartes, Sorbonne Paris Cité, Paris, France. Electronic address: sebastien.storck@inserm.fr.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Storck
mesh_terms: D000409:Alanine / Q000378:metabolism; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism*; D020134:Catalytic Domain; D002460:Cell Line; D002470:Cell Survival / Q000528:radiation effects; D017124:Conserved Sequence; D004268:DNA-Binding Proteins / Q000172:deficiency / Q000235:genetics*; D004259:DNA-Directed DNA Polymerase / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D004622:Embryo, Mammalian; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D007136:Immunoglobulins / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D009154:Mutation; D010641:Phenotype; D014466:Ultraviolet Rays
chemical_list: D000409:Alanine; D001224:Aspartic Acid; D004268:DNA-Binding Proteins; D004259:DNA-Directed DNA Polymerase; D007136:Immunoglobulins; C425929:DNA polymerase zeta; C486862:REV3L protein, mouse
keywords: DNA damage; DNA polymerase zeta; Rev3l; Somatic hypermutation; Translesion