A mouse PRMT1 null allele defines an essential role for arginine methylation in genome maintenance and cell proliferation.
Protein arginine methyltransferase 1 (PRMT1) is the major enzyme that generates monomethylarginine and asymmetrical dimethylarginine. We report here a conditional null allele of PRMT1 in mice and that the loss of PRMT1 expression leads to embryonic lethality. Using the Cre/lox-conditional system, we show that the loss of PRMT1 in mouse embryonic fibroblasts (MEFs) leads to the loss of arginine methylation of substrates harboring a glycine-arginine rich motif, including Sam68 and MRE11. The loss of PRMT1 in MEFs leads to spontaneous DNA damage, cell cycle progression delay, checkpoint defects, aneuploidy, and polyploidy. We show using a 4-hydroxytamoxifen-inducible Cre that the loss of PRMT1 in MEFs leads to a higher incidence of chromosome losses, gains, structural rearrangements, and polyploidy, as documented by spectral karyotyping. Using PRMT1 small interfering RNA in U2OS cells, we further show that PRMT1-deficient cells are hypersensitive to the DNA damaging agent etoposide and exhibit a defect in the recruitment of the homologous recombination RAD51 recombinase to DNA damage foci. Taken together, these data show that PRMT1 is required for genome integrity and cell proliferation. Our findings also suggest that arginine methylation by PRMT1 is a key posttranslational modification in the DNA damage response pathway in proliferating mammalian cells.
Yu
Zhenbao
Z
Bloomfield Center for Research on Aging, Segal Cancer Centre, Lady Davis Institute for Medical Research, Sir Mortimer B. Davis Jewish General Hospital, Montréal, Québec, Canada.
Chen
Taiping
T
Hébert
Josée
J
Li
En
E
Richard
Stéphane
S
D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D000483:Alleles; D000818:Animals; D001120:Arginine / Q000378:metabolism*; D049109:Cell Proliferation / Q000187:drug effects / Q000528:radiation effects; D004247:DNA / Q000378:metabolism; D004249:DNA Damage; D045643:DNA Repair Enzymes / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects / Q000528:radiation effects; D004622:Embryo, Mammalian / Q000166:cytology; D005047:Etoposide / Q000494:pharmacology; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000201:enzymology / Q000528:radiation effects; D016678:Genome / Q000235:genetics*; D042822:Genomic Instability / Q000187:drug effects / Q000528:radiation effects; D007399:Interphase / Q000187:drug effects / Q000528:radiation effects; D000076228:MRE11 Homologue Protein; D008745:Methylation / Q000187:drug effects / Q000528:radiation effects; D051379:Mice; D008810:Mice, Inbred C57BL; D008938:Mitosis / Q000187:drug effects / Q000528:radiation effects; D011484:Protein-Arginine N-Methyltransferases / Q000235:genetics*; D016601:RNA-Binding Proteins / Q000378:metabolism; D051135:Rad51 Recombinase / Q000378:metabolism; D011839:Radiation, Ionizing
D048868:Adaptor Proteins, Signal Transducing; D004268:DNA-Binding Proteins; C513253:Khdrbs1 protein, mouse; C512689:Mre11a protein, mouse; D016601:RNA-Binding Proteins; D005047:Etoposide; D004247:DNA; D001120:Arginine; D011484:Protein-Arginine N-Methyltransferases; D051135:Rad51 Recombinase; C495154:Rad51 protein, mouse; D000076228:MRE11 Homologue Protein; D045643:DNA Repair Enzymes