The Gpn3 Q279* cancer-associated mutant inhibits Gpn1 nuclear export and is deficient in RNA polymerase II nuclear targeting
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Gpn3 is required for RNA polymerase II (RNAPII) nuclear targeting. Here, we investigated the effect of a cancer-associated Q279* nonsense mutation in Gpn3 cellular function. Employing RNAi, we replaced endogenous Gpn3 by wt or Q279* RNAi-resistant Gpn3R in epithelial model cells. RNAPII nuclear accumulation and transcriptional activity were markedly decreased in cells expressing only Gpn3R Q279*. Wild-type Gpn3R localized to the cytoplasm but a fraction of Gpn3R Q279* entered the cell nucleus and inhibited Gpn1-EYFP nuclear export. This property and the transcriptional deficit in Gpn3R Q279*-expressing cells required a PDZ-binding motif generated by the Q279* mutation. We conclude that an acquired PDZ-binding motif in Gpn3 Q279* caused Gpn3 nuclear entry, and inhibited Gpn1 nuclear export and Gpn3-mediated RNAPII nuclear targeting. © 2017 Federation of European Biochemical Societies.
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colon and skin cancer; Gpn1 nuclear export; Gpn3; PDZ-binding motif Gpn1 protein; Gpn3 protein; guanosine triphosphatase; messenger RNA; RNA polymerase II; unclassified drug; GPN1 protein, human; guanine nucleotide binding protein; guanosine triphosphatase; Parcs protein, human; RNA polymerase II; tumor protein; cell nucleus; cellular distribution; colon cancer; controlled study; cytoplasm; HEK293T cell line; human; human cell; Letter; MCF-12A cell line; nonsense mutation; nuclear export; PDZ domain; priority journal; RNA interference; skin cancer; transcription regulation; breast tumor; enzymology; female; genetics; HEK293 cell line; metabolism; nucleocytoplasmic transport; pathology; stop codon; tumor cell line; Active Transport, Cell Nucleus; Breast Neoplasms; Cell Line, Tumor; Cell Nucleus; Codon, Nonsense; Cytoplasm; Female; GTP Phosphohydrolases; GTP-Binding Proteins; HEK293 Cells; Humans; Neoplasm Proteins; PDZ Domains; RNA Polymerase II
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