TY - JOUR
T1 - A single aspartate mutation in the conserved catalytic site of Rev3L generates a hypomorphic phenotype in vivo and in vitro
AU - Fritzen, Rémi
AU - Delbos, Frédéric
AU - De Smet, Annie
AU - Palancade, Benoît
AU - Canman, Christine E
AU - Aoufouchi, Said
AU - Weill, Jean-Claude
AU - Reynaud, Claude-Agnès
AU - Storck, Sébastien
N1 - This work was supported by the Ligue Nationale contre le Cancer (“Equipe labellisée”), and the Fondation Princesse Grace, and was inititated with an ANR grant “Mutig” (ANR-06-BLAN-082).
PY - 2016/10
Y1 - 2016/10
N2 - 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.
AB - 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.
KW - DNA damage
KW - Rev3I
KW - DNA polymerase zeta
KW - Translesion
KW - Somatic hypermutation
UR - http://www.sciencedirect.com/science/article/pii/S1568786416301616#appd002
U2 - 10.1016/j.dnarep.2016.07.002
DO - 10.1016/j.dnarep.2016.07.002
M3 - Article
C2 - 27481099
SN - 1568-7864
VL - 46
SP - 37
EP - 46
JO - DNA Repair
JF - DNA Repair
ER -