Cell-cycle-specific cDNAs from mammalian cells temperature sensitive for growth

R. R. Hirschhorn, P. Aller, Z. A. Yuan, C. W. Gibson, R. Baserga

Research output: Contribution to journalArticle

Abstract

A library of double-stranded cDNA was constructed from ts13 cells, a G1-specific temperature-sensitive hamster cell line. The cDNAs, cloned into pBR322, were prepared from poly(A)+ mRNA isolated from ts13 cells 6 hr after serum stimulation at the permissive temperature of 34° C. Differential screening of the library with G1-specific and G0-specific single-stranded cDNA probes prepared from the same cells identified 5 cDNA clones whose sequences were preferentially expressed in G1. Levels of RNA complementary to these clones were 3- to 6-fold higher in G1 than in other phases of the cell cycle. When ts13 cells were grafted in G1 at the restrictive temperature of 39.6°C, the levels of RNA complementary to p13-2A9 and p13-4F1 were as high as 10 times that found in a resting population, while the expression of sequences complementary to p13-2A8 did not significantly change from levels found in G0. RNA and Southern gel blot analysis suggest that these cell-cycle-specific clones represent either low copy or moderately repetitive gene sequences. Results with another ts mutant of the cell cycle, tsAF8, which is a ts mutant of RNA polymerase II, showed that these cell-cycle-specific sequences have a rapid turnover. The use of G1-specific ts mutants of the cell cycle provides an approach to determine which cell-cycle-dependent genes are most relevant to cell-cycle progression.

Original languageEnglish (US)
Pages (from-to)6004-6008
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume81
Issue number19 I
DOIs
StatePublished - Dec 1 1984

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