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M E Gottesman

Publications and source records attributed to M E Gottesman.

At least 91 records · Page 5Linked to original sources

Escherichia coli gal operon proteins made after prophage lambda induction.

Expression of the EScherichia coli gal operon under the control of the prophage lambda promoter pL leads to gross discoordinacy of gal expression. Expression of the most promoter-distal cistron galK is much greater than expression of the promoter-proximal cistron galE. We had previously shown that transcription of the gal operon is coordinate after prophage induction. A survey of protein synthesized after prophage induction indicated that lack of expression of galE is due to a failure of translation of the galE sequence in the pL-gal transcript. This failure of translation of the galE sequence may be due to extensive dyad symmetry present in the vicinity of the gal promoter region of the pL-gal transcript. This symmetry could result in a ribonucleic acid stem-loop structure, blocking the attachment of ribosomes at the Shine-Dalgarno sequence of galE. To test this model, strains bearing the IS1 or IS2 insertion, deletion, or new promoter mutation within the symmetrical region were constructed. The restoration of some galE expression after such disruptions of the symmetrical region indicated that the ribonucleic acid stem-loop structure did play a role in the discoordinate expression of gal from pL. However, failure to obtain galE expression coordinated with high levels of galK expression suggested that other components were involved, perhaps other symmetries between galE and the pL transcript.

Bacteriophage lambda↗

In vitro repression of the transcription of gas operon by purified gal repressor.

We have studied the in vitro repression of gal mRNA synthesis by the gal repressor from Escherichia coli. By use of a four-step purification procedure involving chromatography on phosphocellulose, DEAE-cellulose, and an affinity resin, the gal repressor has been purified about 1600-fold from a crude cell extract. The purification was aided by use of a cell extract made after prophage induction of cells lysogenic for bacteriophage lambda that carries the gal repressor gene (galR). The highly purified gal repressor is an effective and specific repressor of in vitro synthesis of gal mRNA with lambda gal DNA as template. Both D-fucose and D-galactose overcome the action of gal repressor; the half-maximal concentrations of D-fucose and D-galactose for overcoming the action of repressor are 1 mM and 0.5 mM, respectively. The repressor fails to repress gal-specific transcription when the gal DNA contains a cis-dominant operator constitutive (O(c)) mutation. We conclude that the gal repressor recognizes the gal operator site and acts by preventing gal transcription.

Ammonium Sulfate↗