A bipartite nuclear localization signal is required for p53 nuclear import regulated by a carboxyl-terminal domain.
Abnormal p53 cellular localization has been considered to be one of the mechanisms that could inactivate p53 function. To understand the regulation of p53 cellular trafficking, we have previously identified two p53 domains involved in its localization. A basic domain, Lys(305)-Arg(306), is required for p53 nuclear import, and a carboxyl-terminal domain, namely the cytoplasmic sequestration domain (CSD) from residues 326-355, could block the nuclear import of Lys(305) or Arg(306) mutated p53. To characterize further the function of these two domains, we demonstrate in this report that the previously described major nuclear localization signal works together with Lys(305)-Arg(306) to form a bipartite and functional nuclear localization sequence (NLS) for p53 nuclear import. The CSD could block the binding of p53 to the NLS receptor, importin alpha, and reduce the efficiency of p53 nuclear import in MCF-7, H1299, and Saos-2 cells. The blocking effect of the CSD is not due to the enhancement of nuclear export or oligomerization of the p53. These results indicate that the CSD can regulate p53 nuclear import by controlling access of the NLS to importin alpha binding.
Liang
S H
SH
Department of Internal Medicine, University of Michigan Medical Center, Ann Arbor, Michigan 48109-0936, USA.
Clarke
M F
MF
D000595:Amino Acid Sequence; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis; D019913:Nuclear Localization Signals / Q000235:genetics*; D009687:Nuclear Proteins / Q000378:metabolism; D011485:Protein Binding / Q000235:genetics; D011487:Protein Conformation; D014162:Transfection; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry* / Q000235:genetics; D028901:alpha Karyopherins
D000952:Antigens, Polyomavirus Transforming; D019913:Nuclear Localization Signals; D009687:Nuclear Proteins; D016159:Tumor Suppressor Protein p53; D028901:alpha Karyopherins