p53 binds the nuclear matrix in normal cells: binding involves the proline-rich domain of p53 and increases following genotoxic stress.
The tumour suppressor p53 is a multifunctional protein important for the maintenance of genomic integrity. It is able to form molecular complexes with different DNA targets and also with cellular proteins involved in DNA transcription and DNA repair. In mammalian cells the biochemical processing of DNA occurs on a nuclear sub-structure termed the nuclear matrix. Previously Deppert and co-workers have identified p53 in association with the nuclear matrix in viral- and non-viral transformed cell lines. In the present study we demonstrate, for the first time, that p53 is bound to the nuclear matrix in primary cultures of normal mammalian cells and that this binding increases following DNA damage. Analysis of cell lines expressing structural mutants of p53 revealed that association with the nuclear matrix is independent of the tertiary and quaternary structure of p53. However, the proline-rich domain towards the N-terminus of p53 (residues 67 to 98) appeared important for binding to the nuclear matrix. This was demonstrated by TET-ON regulated expression of p53-derived constructs in p53(-/-) murine embryonic fibroblasts (MEF p53(-/-)). The proline-rich domain of p53 has potential for SH3 protein-protein interaction, and has a role in p53-mediated apoptosis and possibly base excision repair of DNA damage. We discuss our observations in relation to the ability of p53 to facilitate DNA repair and also review evidence indicating that matrix-bound p53 in SV40-transformed cells may facilitate the transforming potential of SV40 large T antigen.
Jiang
M
M
YCR P53 Research Group, Department of Biology, University of York, YO10 5DD, UK.
Axe
T
T
Holgate
R
R
Rubbi
C P
CP
Okorokov
A L
AL
Mee
T
T
Milner
J
J
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000502:physiology; D002453:Cell Cycle; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D004249:DNA Damage; D017353:Gene Deletion; D006801:Humans; D051379:Mice; D009154:Mutation; D015530:Nuclear Matrix / Q000378:metabolism*; D011392:Proline / Q000737:chemistry; D013312:Stress, Physiological; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D000952:Antigens, Polyomavirus Transforming; D016159:Tumor Suppressor Protein p53; D011392:Proline