p53 in cytoplasm exerts 3'→5' exonuclease activity with dsRNA.
Double-stranded RNA (dsRNA) is a biologically active molecule that plays important roles in normal cell growth and function. Accordingly, the cell uses multiple mechanisms to control its level. The tumor suppressor protein p53 possesses intrinsic 3'→5' exonuclease activity. The aim of the present study was to elucidate the degradation of dsRNA by the exonuclease activity of p53. The results show that recombinant, purified wtp53 and endogenous protein in cytoplasmic fractions of cells remove nucleotides from 3'-ends of dsRNA. Several lines of evidence support a connection between p53 and dsRNase activity in cytoplasm: (1) this activity parallels the status of endogenous cytoplasmic p53; (2) the endogenous exonuclease displays a similar dsRNA excision profile characteristic for purified wtp53; (3) cytoplasmic fractions of HCT116(p53+/+) cells exert higher levels of exonuclease activity compared to those of HCT116(p53-/-) cells; (4) transfection of the wtp53, but not exonuclease-deficient mutant p53-R175H, into HCT116 (p53-/-) cells induced high levels of dsRNase activity in cytoplasm; (5) the accumulation of p53 in cytoplasm following the γ-irradiation stress stimuli correlates with the increase in the excision of dsRNA and (6) the dsRNA forms a complex with a protein that can be disrupted by an anti-p53 antibody. Our data suggest that the degradation of dsRNA by p53 protein may direct either the complete degradation of and decrease in the level of dsRNA or incomplete degradation and the generation of short dsRNA products. The possible roles of p53 dsRNase activity in cytoplasm in the inhibition of translation and induction of cell apoptosis, is discussed.
Infectious Diseases Unit, Sheba Medical Center, Tel Hashomer, Ramat Gan, Israel.
Shai
S
Grinberg
Gabriel
G
Teiblum
Galia
G
Rahav
Mary
M
Bakhanashvili
D000911:Antibodies, Monoclonal; D017209:Apoptosis; D015153:Blotting, Western; D045744:Cell Line, Tumor; D003593:Cytoplasm / Q000201:enzymology*; D004249:DNA Damage; D024202:Electrophoretic Mobility Shift Assay; D005092:Exonucleases / Q000378:metabolism*; D006801:Humans; D012330:RNA, Double-Stranded / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D012330:RNA, Double-Stranded; D011994:Recombinant Proteins; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D005092:Exonucleases