Cookies on this website
We use cookies to ensure that we give you the best experience on our website. If you click 'Continue' we'll assume that you are happy to receive all cookies and you won't see this message again. Click 'Find out more' for information on how to change your cookie settings.

The p16INK4A cyclin-dependent kinase (Cdk) inhibitor is now recognized as a major tumor suppressor that is inactivated by a variety of mechanisms in a wide range of human cancers. It is also implicated in the mechanisms underlying replicative senescence since p16INK4A RNA and protein accumulate as cells approach their proscribed limit of population doublings in tissue culture. To obtain further evidence of its role in senescence, we have sought ways of overexpressing p16INK4A in primary human diploid fibroblasts (HDF). To circumvent the low transfection efficiency of primary cells we have exploited a recombinant form of the full-length p16INK4A protein fused to a 16 amino acid peptide from the Drosophila antennapedia protein. This peptide has the capacity to cross both cytoplasmic and nuclear membranes allowing the direct introduction of the active protein to primary cells. Here, we show that antennapedia-tagged wild-type p16INK4A protein, but not a functionally compromised tumor-specific variant, causes G1 arrest in early passage HDFs by inhibiting the phosphorylation of the retinoblastoma protein. Significantly, the arrested cells display several phenotypic features that are considered characteristic of senescent cells. These data support a role for p16INK4A in replicative senescence and raise the possibility of using the antennapedia-tagged protein therapeutically.

Original publication

DOI

10.1016/s0014-5793(98)00426-8

Type

Journal article

Journal

FEBS Lett

Publication Date

08/05/1998

Volume

427

Pages

203 - 208

Keywords

Antennapedia Homeodomain Protein, Cell Division, Cell Nucleus, Cells, Cultured, Cellular Senescence, Cyclin-Dependent Kinase Inhibitor p16, Diploidy, Escherichia coli, Fibroblasts, G1 Phase, Homeodomain Proteins, Humans, Nuclear Proteins, Phenotype, Phosphorylation, Recombinant Fusion Proteins, Retinoblastoma Protein, Transcription Factors