D-type cyclin expression is decreased and p21 and p27 CDK inhibitor expression is increased when tsBN462 CCG1/TAF11250 mutant cells arrest in G1 at the restrictive temperature

Takeshi Sekiguchi, Eishi Noguchi, Toshiro Hayashida, Torahiko Nakashima, Hideo Toyoshima, Takeharu Nishimoto, Tony Hunter

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Background: The tsBN462 temperature-sensitive mutant hamster cell line exhibits cell cycle arrest and apoptosis at the restrictive temperature of 39.5 °C, due to a point mutation in the CCG1/ TAF11250 gene, which encodes a component of the general transcription factor TFIID. Results: We now report that CCG1/TAFII250 persisted as a complex with TBP and associated proteins (TAFs) in tsBN462 cells at the restrictive temperature. FACScan analysis revealed that the tsBN462 mutation resulted in a failure to progress out of G0 into G1. Using two-dimensional gel electrophoresis we observed a decrease in the synthesis of several proteins, starting in the middle of the G1 phase, becoming very pronounced during late G1. The expression of the immediate early genes c-fos, c-jun and c-myc was normally induced by seruin treatment of quiescent cells at the restrictive temperature, whereas expression of cyclins A, Dl and D3 was reduced. Expression of the cyclin-dependent kinase (CDK) inhibitor proteins p21 and p27 was enhanced. Consistent with the decreased cyclin D and increased p21/p27 expression, we found that phosphorylation of Rb was decreased at 39.5 °C. Cyclin A-, E- and Cdk2-associated historic HI kinase activity was reduced concomitantly with the increase in p21 protein. Conclusion: Decreased cyclin/Cdk kinase activity and decreased Rb phosphorylation are possible causes of G1 cell cycle arrest in tsBN462 cells at the restrictive temperature.

Original languageEnglish
Pages (from-to)687-705
Number of pages19
JournalGenes to Cells
Volume1
Issue number7
DOIs
Publication statusPublished - 1996

All Science Journal Classification (ASJC) codes

  • Genetics
  • Cell Biology

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