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Decreased levels of collagen mRNA in rous sarcoma virus-transformed chick embryo fibroblasts.

Authors
  • Howard, B H
  • Adams, S L
  • Sobel, M E
  • Pastan, I
  • de Crombrugghe, B
Type
Published Article
Journal
The Journal of biological chemistry
Publication Date
Aug 25, 1978
Volume
253
Issue
16
Pages
5869–5874
Identifiers
PMID: 78927
Source
Medline
License
Unknown

Abstract

We have found that double-stranded cDNA synthesized in extended reactions by avian myeloblastosis virus reverse transcriptase is suitable substrate for a variety of restriction endonucleases. Experiments in which rabbit reticulocyte mRNA was reverse-transcribed and restricted to generate beta-globin-specifihe nucleotide sequence of beta-globin mRNA. This method has been applied to study collagen mRNA synthesis in normal and Rous sarcoma virus (RSV)-transformed chick embryo fibroblasts. Characteristic sets of collagen cDNA restriction fragments were produced from RNA fractions rich in collagen message activity. These sets of cDNA fragments, generated by the restriction endonucleases Hae III and Hap II, provided a convenient marker for the presence of collagen mRNA sequences. Equal quantities of high molecular weight mRNA from chick embryo fibroblasts (CEF) and RSV-CEF were reverse-transcribed and the resulting cDNA was restricted. The relative yields of collagen cDNA fragments from such reactions strongly suggest that the decrease in functional collagen RNA following RSV-induced transformation of CEF represents a decrease in the copy number of collagen messenger sequences. The potential of this approach for the study of regulation in other systems is discussed.

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