JBC Invitrogen Ultrasensitive Cytokine Assays

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Regulation of Lac Transcription in Escherichia coli by Cyclic Adenosine 3',5'-Monophosphate

STUDIES WITH DEOXYRIBONUCLEIC ACID-RIBONUCLEIC ACID HYBRIDIZATION AND HYBRIDIZATION COMPETITION

Harold E. Varmus 1, Robert L. Perlman 1, and Ira Pastan 1

From the 1 From the Clinical Endocrinology Branch, National Institute of Arthritis and Metabolic Diseases and Laboratory of Molecular Biology, National Cancer Institute, Bethesda, Maryland 20014

Cyclic adenosine 3',5'-monophosphate (cyclic AMP) is required for the synthesis of lac mRNA and ß-galactosidase in Escherichia coli and cell-free extracts. Competition hybridization experiments, described in this report, show that the concentrations of lac mRNA in E. coli are controlled by cyclic AMP, as well as by inducer. Both transient and permanent repression by glucose affect lac mRNA production and are reversed by cyclic AMP. In an adenyl cyclase-deficient mutant strain, lac mRNA is not synthesized until the cells are supplied with cyclic AMP. A mutant strain lacking a recently described "cyclic AMP receptor protein" does not make lac mRNA despite exogenous cyclic AMP. We conclude that cyclic AMP, cyclic AMP receptor protein, an intact promoter locus, and other unidentified factor or factors are necessary for efficient transcription of the lac operon.

Submitted on June 5, 1970




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