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The C-terminal Region of p21 Is Involved in Proliferating Cell Nuclear Antigen Binding but Does Not Appear to Be Required for Growth Inhibition

Nakanishi, Makoto ; Robetorye, Ryan S. ; Pereira-Smith, Olivia M. ; Smith, James R.

The Journal of biological chemistry, 1995-07, Vol.270 (29), p.17060-17063 [Periódico revisado por pares]

American Society for Biochemistry and Molecular Biology

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  • Título:
    The C-terminal Region of p21 Is Involved in Proliferating Cell Nuclear Antigen Binding but Does Not Appear to Be Required for Growth Inhibition
  • Autor: Nakanishi, Makoto ; Robetorye, Ryan S. ; Pereira-Smith, Olivia M. ; Smith, James R.
  • É parte de: The Journal of biological chemistry, 1995-07, Vol.270 (29), p.17060-17063
  • Descrição: The cyclin-dependent kinase (Cdk) inhibitor p21 has been found to be involved in cell senescence, cell cycle arrest, and differentiation. p21 inhibits the activity of several Cdks, in contrast to other inhibitors such as p15 and p16 , which act on specific cyclin-Cdk complexes. Of interest were reports that p21 also bound proliferating cell nuclear antigen (PCNA), an auxiliary protein for DNA polymerase , and inhibited DNA replication but not DNA repair in vitro . To better understand the function of this interaction in vivo , we first determined the region of p21 that was needed for PCNA binding. Analysis of deletion mutants of p21 , which covered the majority of the protein, revealed that deletion of either amino acids 142-147 or 149-154 resulted in loss of ability to bind a glutathione S -transferase-PCNA fusion protein. Site-directed mutagenesis in this region led to the identification of the PCNA binding motif RQ XX MT X FY XXX R and demonstrated that mutation of either amino acid Met-147 or Phe-150 resulted in almost complete ablation of PCNA binding. Interestingly, when we determined DNA synthesis inhibitory activity of deletion mutants or point mutants that were unable to bind Cdk2 and/or PCNA, we found that loss of binding to PCNA did not affect inhibitory activity, whereas lack of Cdk2 binding greatly reduced the same. This result suggests that the primary mechanism for inhibition of DNA synthesis by p21 occurs via inhibition of Cdk activity.
  • Editor: American Society for Biochemistry and Molecular Biology
  • Idioma: Inglês
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  • Realização: ABCD-USP USP   Logos de Redes Sociais: Freepik

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