Expression of a gene for mouse eucaryotic elongation factor Tu during murine erythroleukemic cell differentiation.

W. W. Roth, P. W. Bragg, M. V. Corrias, N. S. Reddy, J. N. Dholakia, A. J. Wahba

Research output: Contribution to journalArticlepeer-review

Abstract

The eucaryotic elongation factor Tu (eEF-Tu) is a single polypeptide with an approximate Mr of 53,000. During protein synthesis eEF-Tu promotes the binding of aminoacyl-tRNA to the ribosome. To study the expression of the gene(s) for this factor, a genomic clone was isolated that contains a mouse eEF-Tu gene. We screened a phage genomic library with a synthetic oligonucleotide probe complementary to a region of the Saccharomyces cerevisiae and Artemia sp. eEF-Tu genes which codes for an area that is highly conserved between both yeast and Artemia sp. eEF-Tu. From approximately 75,000 phage plaques we obtained five isolates with apparently identical inserts. All five clones contained a 3.8-kilobase EcoRI fragment that hybridized to additional oligonucleotide probes corresponding to different conserved regions of eEF-Tu. We sequenced the 5' end of one genomic clone and determined the length of the cloned fragment that was protected by eEF-Tu mRNA in S1 nuclease protection assays. A quantitative S1 nuclease protection assay was used to compare the relative steady-state levels of eEF-Tu mRNA in total mRNA in total RNA isolated from hexamethylene-bisacetamide-induced murine erythroleukemia cells. The results show a dramatic reduction in the steady-state level of eEF-Tu mRNA as differentiation proceeds. A similar reduction in transcription of eEF-Tu mRNA was observed in isolated nuclei. Finally, we examined the in vivo synthesis of eEF-Tu during differentiation and found that it declined in a manner parallel to the decline in the steady-state level of eEF-Tu mRNA.(ABSTRACT TRUNCATED AT 250 WORDS)

Original languageEnglish
Pages (from-to)3929-3936
Number of pages8
JournalMolecular and Cellular Biology
Volume7
Issue number11
Publication statusPublished - Nov 1987

ASJC Scopus subject areas

  • Cell Biology
  • Genetics
  • Molecular Biology

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