Differential protein expression, DNA binding and interaction with SV40 large tumour antigen implicate the p63-family of proteins in replicative senescence.
P53 activity plays a key role in mammalian cells when they undergo replicative senescence at their Hayflick limit. To determine whether p63 proteins, members of the family of p53-related genes, are also involved in this process, we examined their expression in serially passaged rat embryo fibroblasts. Upon senescence, two truncated DeltaNp63 proteins decreased in abundance whereas two TAp63 isoforms accumulated. 2-D gel analysis showed that the DeltaNp63 proteins underwent post-translational modifications in both proliferating and senescent cells. Direct binding of DeltaNp63 proteins to a p53 consensus motif was greater in proliferating cells than senescent cells. In contrast p63alpha isoforms bound to DNA in a p53 dependent manner and this was higher in senescent cells than proliferating cells. An interaction of p63alpha proteins with SV40 large tumour antigen was also detected and ectopic expression of DeltaNp63alpha can extend the lifespan of rat embryo fibroblasts. Taken together the results indicate that p63 proteins may play a role in replicative senescence either by competition for p53 DNA binding sites or by direct interaction with p53 protein bound to DNA.
Djelloul
Siham
S
Ludwig Institute for Cancer Research, Royal Free and University College School of Medicine, Courtauld Building, 91 Riding House Street, London W1W 7BS, UK.
Tarunina
Marina
M
Barnouin
Karin
K
Mackay
Alan
A
Jat
Parmjit S
PS
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000378:metabolism*; D001665:Binding Sites; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D016922:Cellular Senescence; D016384:Consensus Sequence; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000032:analysis / Q000276:immunology; D015180:Electrophoresis, Gel, Two-Dimensional; D024202:Electrophoretic Mobility Shift Assay; D005347:Fibroblasts / Q000502:physiology; D016158:Genes, p53; D008565:Membrane Proteins; D019255:NADPH Oxidases; D010750:Phosphoproteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism* / Q000502:physiology*; D020033:Protein Isoforms / Q000276:immunology / Q000378:metabolism / Q000502:physiology; D011499:Protein Processing, Post-Translational; D011506:Proteins; D051381:Rats; D017207:Rats, Sprague-Dawley; D017384:Sequence Deletion; D015534:Trans-Activators / Q000235:genetics / Q000276:immunology / Q000378:metabolism* / Q000502:physiology*; D016159:Tumor Suppressor Protein p53
D000952:Antigens, Polyomavirus Transforming; C081876:CKAP4 protein, human; D004268:DNA-Binding Proteins; D008565:Membrane Proteins; D010750:Phosphoproteins; D020033:Protein Isoforms; D011506:Proteins; C108581:Tp63 protein, rat; D015534:Trans-Activators; D016159:Tumor Suppressor Protein p53; D004247:DNA; D019255:NADPH Oxidases; C113333:NCF4 protein, human