Identification of new drug sensitivity genes using genetic suppressor elements: protein arginine N-methyltransferase mediates cell sensitivity to DNA-damaging agents.
Genetic suppressor elements (GSEs) are cDNA fragments encoding either truncated proteins, acting as dominant-negative mutants, or inhibitory antisense RNA segments counteracting with the gene from which they are derived. To identify genes controlling the cell response to cytotoxic agents, a normalized retroviral library of randomly fragmented cDNAs from Chinese hamster cell line DC-3F was screened for GSEs conferring resistance to the topoisomerase II inhibitor 9-OH-ellipticine. From 218 cDNA fragments isolated, 11 functional GSEs, corresponding to at least 8 independent genes, were selected. The gene corresponding to the most abundant GSE encodes two proteins, p77 and p82, highly homologous to proteins detected in various species and carrying the sequence motifs characteristic of the protein arginine N-methyltransferase family. Furthermore, a methylase activity was observed on myelin basic protein in immunoprecipitates of hemagglutinin-tagged p77 and p82. Therefore, p77 and p82 are the first identified members of a new protein arginine N-methyltransferase family. A decreased expression of these enzymes is associated with either resistance or hypersensitivity to a broad range of DNA-damaging agents. Our data indicate that down-regulation of these enzymes in the GSE-expressing cells would alter one or several steps downstream of the drug-target interaction in the drug-response pathway.
Gros
Laurent
L
Centre National de la Recherche Scientifique UMR 8532, Institut Gustave Roussy, 94805 Villejuif, France.
Delaporte
Charlotte
C
Frey
Stéphane
S
Decesse
Julien
J
de Saint-Vincent
Bruno Robert
BR
Cavarec
Laurent
L
Dubart
Anne
A
Gudkov
Andrei V
AV
Jacquemin-Sablon
Alain
A
D000595:Amino Acid Sequence; D000964:Antimetabolites, Antineoplastic / Q000633:toxicity*; D002166:Camptothecin / Q000633:toxicity; D002455:Cell Division / Q000187:drug effects; D004249:DNA Damage; D004305:Dose-Response Relationship, Drug; D019008:Drug Resistance, Neoplasm; D004354:Drug Screening Assays, Antitumor; D004611:Ellipticines / Q000633:toxicity*; D005047:Etoposide / Q000633:toxicity; D006801:Humans; D008969:Molecular Sequence Data; D011484:Protein-Arginine N-Methyltransferases / Q000235:genetics* / Q000378:metabolism*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013489:Suppression, Genetic; D014407:Tumor Cells, Cultured
D000964:Antimetabolites, Antineoplastic; D004611:Ellipticines; D005047:Etoposide; C008009:9-hydroxyellipticine; D011484:Protein-Arginine N-Methyltransferases; D002166:Camptothecin