a monoclonal antibody against p53 cross-reacts with processing bodies对p53单克隆抗体交叉反应处理尸体.pdfVIP

A Monoclonal Antibody against p53 Cross-Reacts with Processing Bodies ´ 1,2 1,2,3 1,2,3 1,2,3 Marıa Gabriela Thomas *, Luciana Luchelli , Malena Pascual , Vanesa Gottifredi , Graciela Lidia Boccaccio1,2,3 1 Instituto Leloir Av. Patricias Argentinas Buenos Aires, Argentina, 2 IIBBA-CONICET, Buenos Aires, Argentina, 3 Facultad de Ciencias Exactas y Naturales, University of Buenos Aires, Argentina Abstract The p53 tumor suppressor protein is an important regulator of cell proliferation and apoptosis. p53 can be found in the nucleus and in the cytosol, and the subcellular location is key to control p53 function. In this work, we found that a widely used monoclonal antibody against p53, termed Pab 1801 (Pan antibody 1801) yields a remarkable punctate signal in the cytoplasm of several cell lines of human origin. Surprisingly, these puncta were also observed in two independent p53-null cell lines. Moreover, the foci stained with the Pab 1801 were present in rat cells, although Pab 1801 recognizes an epitope that is not conserved in rodent p53. In contrast, the Pab 1801 nuclear staining corresponded to genuine p53, as it was upregulated by p53-stimulating drugs and absent in p53-null cells. We identified the Pab 1801 cytoplasmic puncta as P Bodies (PBs), which are involved in mRNA regulation. We found that, in several cell lines, including U2OS, WI38, SK-N-SH and HCT116, the Pab 1801 puncta strictly colocalize with PBs identified with specific antibodies against the PB components Hedls, Dcp1a, Xrn1 or Rck/p54.