Journal ArticleDOI
Purification and properties of the sigma subunit of Escherichia coli DNA-dependent RNA polymerase.
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TLDR
Denaturation--renaturation studies indicate that sigma is capable of an unusually rapid and complete recovery of activity after being subjected to denaturing conditions.Abstract:
An improved purification procedure is described for the sigma subunit of escherichia coli DNA-dependent RNA polymerase [ribonucleoside triphosphate:RNA nucleotidyl-transferase, EC 2.7.7.6]. The method involves chromatography of purified RNA polymerase on single-stranded DNA-agarose, Bio-Rex 70, and finally Ultragel AcA44. The sigma factor obtained is electrophoretically pure with a yield of about 40%. A number of the chemical--physical properties of sigma are presented. A molecular weight of 82,000 was determined by phosphate buffered sodium dodecyl sulfate--polyacrylamide gel electrophoresis. Ultraviolet absorption spectra were used to determine an E280nm 1% of 8.4. The amino acid composition and 12-residue N-terminal sequence (Met-Glx-Glx-Asx-Pro-Glx-(Ser or Cys)-Glx-Leu-Lys-Leu-Leu) of sigma have been determined. The isoelectric focusing properties of sigma are presented. Denaturation--renaturation studies indicate that sigma is capable of an unusually rapid and complete recovery of activity after being subjected to denaturing conditions. A stable, 40,000-dalton fragment is generated from sigma by mild trypsin treatment.read more
Citations
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Journal ArticleDOI
A gel electrophoresis method for quantifying the binding of proteins to specific DNA regions: application to components of the Escherichia coli lactose operon regulatory system
Mark M. Garner,Arnold Revzin +1 more
TL;DR: It is demonstrated that even when pre-formed in the presence of CAP-cAMP, the polymerase-promoter open complex becomes unstable if CAP is then selectively removed, and this gel method is applied to the study of the E. coli lactose operon regulatory system.
Journal ArticleDOI
Elution of proteins from sodium dodecyl sulfate-polyacrylamide gels, removal of sodium dodecyl sulfate, and renaturation of enzymatic activity: Results with sigma subunit of Escherichia coli RNA polymerase, wheat germ DNA topoisomerase, and other enzymes
TL;DR: It is shown how the method can be used to determine the approximate molecular weight of the DNA topoisomerase polypeptide by sectioning a gel on which a partially pure sample has been fractionated by electrophoresis.
Journal ArticleDOI
Multiple sigma subunits and the partitioning of bacterial transcription space.
Tanja M. Gruber,Carol A. Gross +1 more
TL;DR: The sigma repertoire of Escherichia coli, Bacillus subtilis, Streptomyces coelicolor, and cyanobacteria is used to illustrate the different strategies utilized to organize transcriptional space using multiple sigma factors.
Journal ArticleDOI
Mapping and sequencing of mutations in the Escherichia coli rpoB gene that lead to rifampicin resistance.
Ding Jun Jin,Carol A. Gross +1 more
TL;DR: A comprehensive analysis of Rifr mutations to identify their structural and functional effects on RNA polymerase and discusses the implications of the results with regards to the structure of the rifampicin binding site.
Journal ArticleDOI
The htpR gene product of E. coli is a sigma factor for heat-shock promoters
TL;DR: HtpR is a sigma factor that promotes transcription initiation at heat-shock promoters and is proposed to be renamed rpoH and that the gene product be called sigma-32.
References
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Journal ArticleDOI
Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
TL;DR: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products.
Journal Article
Cleavage of structural proteins during the assemble of the head of bacterio-phage T4
TL;DR: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products as mentioned in this paper.
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High resolution two-dimensional electrophoresis of proteins.
TL;DR: This technique provides a method for estimation of the number of proteins made by any biological system and can resolve proteins differing in a single charge and consequently can be used in the analysis of in vivo modifications resulting in a change in charge.
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Spectroscopic determination of tryptophan and tyrosine in proteins.
TL;DR: A procedure is presented which strongly reduces or elimi- nates these interactions, normalizes their absorption, and consequently permits a more precise analysis of tryptophan and tyrosine in proteins.
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On the determination of cystine as cysteic acid.
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