The T197A Knock-in Model of Cdkn1b Gene to Study the Effects of p27 Restoration In Vivo.

Abstract

The CDK inhibitor, p27kip1, encoded by the Cdkn1b gene can negatively modulate cell proliferation. The control of p27 activity during the cell cycle is regulated at multiple levels, including transcription, translation, and protein stability. The last residue of p27 (threonine 198 in human, threonine 197 in mouse) is involved in the control of protein stability. We have generated a murine knock-in model (Cdkn1b T197A) in which threonine 197 is replaced by alanine, which renders p27 protein highly unstable due to a high rate of proteasomal degradation. Expectedly, Cdkn1b T197A/T197A mice present with increased body size and weight, organomegaly, and multiple organ hyperplasia, similar to what is observed in Cdkn1b KO/KO mice. We investigated the effects exerted by the restoration of normal levels of p27 protein in the tissue of Cdkn1b T197A/T197A mice. We found that proteasome inhibition with bortezomib rescues the hyperplasia induced by the lack of p27 expression in Cdkn1b T197A/T197A but not in Cdkn1b KO/KO mice. However, BAY 11-7082, a proteasome inhibitor that stabilizes IκB but not p27, fails to rescue hyperplasia in Cdkn1b T197A/T197A mice. Bortezomib increases p27 half-life and reduces the proliferation in MEFs derived from Cdkn1b T197A/T197A but not from Cdkn1b WT/WT mice, whereas BAY 11-7082 had no effect on the protein levels of p27 and on the proliferation rate of Cdkn1b T197A/T197A MEFs.The results presented here demonstrate that Cdkn1b T197A/T197A mice represent an attractive in vivo model to investigate whether the targeting of p27 degradation machinery might prove beneficial in the treatment of a variety of human proliferative disorders caused by increased turnover of p27 protein.

• Authors: Carmela De Marco, Nicola Rinaldo, Fernanda De Vita, Floriana Forzati, Elvira Caira, Valentina Iovane, Orlando Paciello, Donatella Montanaro, Sara D'Andrea, Gustavo Baldassarre, Serenella Papparella, Donatella Malanga, Alfonso Baldi, Giuseppe Viglietto
• Journal: Molecular cancer therapeutics (Mol Cancer Ther) Vol. 18 Issue 2 Pg. 482-493 (02 2019) ISSN: 1538-8514 [Electronic] United States
• PMID: 30425132 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: ©2018 American Association for Cancer Research.

Chemical References

• 3-(4-methylphenylsulfonyl)-2-propenenitrile
• Cdkn1b protein, mouse
• Nitriles
• Sulfones
• Cyclin-Dependent Kinase Inhibitor p27
• Bortezomib
• Proteasome Endopeptidase Complex

Topics

• Amino Acid Substitution
• Animals
• Bortezomib (pharmacology)
• Cyclin-Dependent Kinase Inhibitor p27 (chemistry, genetics)
• Gene Knock-In Techniques
• Hyperplasia
• Mice
• Models, Animal
• Nitriles (pharmacology)
• Proteasome Endopeptidase Complex (metabolism)
• Protein Stability
• Proteolysis
• Sulfones (pharmacology)