Cloned DNA fragment specifying major outer membrane protein a in Escherichia coli K-12

Gayda, Randall C.; Markovitz, Alvin

doi:10.1128/jb.136.1.369-380.1978

Cited by 35 publications

(15 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It has been suggested that protein a is revolved in repression of capsular polysaccharide synthesis [20,21 ]. The strains lacking a which were described here, however, were normal with respect to mucoidy; that is, when grown on minimal plates at 30°C or 37°C, they formed mucoid colonies only at 30°C.…”

Section: Discussionmentioning

confidence: 99%

Escherichia coli K-12 mutants that lack major outer membrane protein a

Earhart¹,

Lundrigan²,

Pickett³

et al. 1979

FEMS Microbiology Letters

View full text Add to dashboard Cite

Section: Discussionmentioning

confidence: 99%

Escherichia coli K-12 mutants that lack major outer membrane protein a

Earhart¹,

Lundrigan²,

Pickett³

et al. 1979

FEMS Microbiology Letters

View full text Add to dashboard Cite

“…Protein labeling and analysis by PAGE-SDS. Plasmidcontaining minicells were isolated and labeled with [35S]methionine as described previously (13,47). Whole cells with or without plasmids were induced with nalidixic acid and labeled with [35S]methionine as follows.…”

Section: Methodsmentioning

confidence: 99%

“…Protein samples and standards were prepared and subjected to PAGE-SDS in one dimension as described previously with approximately equal quantities of total radioactivity in each gel slot (13,47). The preparation of cells for twodimensional gel electrophoresis has been described previously (46), as have the details for the two-dimensional gel electrophoresis used here (3,(41)(42)(43).…”

Section: Methodsmentioning

confidence: 99%

“…Cell fractionation procedures. Preparation of whole cell extracts, isolation of Sarkosyl-insoluble membranes, and separation of inner and outer membranes by isopycnic sucrose gradients were as described previously (13). Membranes layered on the isopycnic gradient were obtained by either French pressure cell lysis at 16,000 lb/in2 (49) or sonic disruption.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Regulation of cell division in Escherichia coli: SOS induction and cellular location of the sulA protein, a key to lon-associated filamentation and death

Schoemaker¹,

Gayda²,

Markovitz³

1984

J Bacteriol

Self Cite

136

View full text Add to dashboard Cite

Mutations in sulA (sfiA) block the filamentation and death of capR (ion) mutants that occur after treatments that either damage DNA or inhibit DNA replication and thereby induce the SOS response. Previous suiA-iacZ gene fusion studies showed that sulA is transcriptionally regulated by the SOS response system (lexAlrecA). SulA protein has been hypothesized to be additionally regulated proteolytically through the capR (lon) protease, i.e., in lon mutants lacking a functional ATP-dependent protease there would be more SulA protein. A hypothesized function for SulA protein is an inhibitor of cell septation. To investigate aspects of this model, we attempted to construct Ion, Ion sulA, and Ion sulB strains containing multicopy plasmids specifying the suiA+ gene. Multicopy suIA+ plasmids could not be established in Ion strains because more SulA protein accumulates than in a Ion' strain. When the sulA gene was mutated by a mini-Mu transposon the plasmid could be established in the Ion strains. In contrast, suiA+ plasmids could be established in lon+, lon sulA, and Ion sulB strains. The suiA+ plasmids caused Ion sulA and Ion sulB cells to exist as filaments without SOS induction and to be sensitive to UV light and nitrofurantoin. Evidence implicated higher basal levels of SulA protein in these Ion plasmid suiA + strains as the cause of filamentation. We confirmed that the SulA protein is an 18-kilodalton polypeptide and demonstrated that it was induced by treatment with nalidixic acid. The SulA protein was rapidly degraded in a lon+ strain, but was comparatively more stable in vivo in a lon sulB mutant. Furthermore, the SulA protein was localized to the membrane by several techniques. enzymatic activities: an ATP hydrolysis-dependent protease activity (9, 10; M. F. Charette, Ph.D. thesis, University of Chicago, 1981), DNA stimulated ATPase activity (7a), and nucleic acid-binding activity (56; Charette, Ph.D. thesis). In addition, a defective CapR protein (capR9 allele) has also been purified. The CapR9 protein has retained the general nucleic acid affinity (9, 56), but has lost the protease activity (8,9). In Ion mutants, second site mutations in sul (suppressor of Ion) prevent the filamentation and UV sensitivity without affecting the mucoid phenotype of the cells (14,17,26,27). These mutants were isolated as UV-, nitro-NF-, or methyl methanesulfonate-resistant derivatives of lon strains. The sul mutations are located at two loci on the E. coli chromosome; sulA is near pyrD (22 min), and sulB is near leu (2 min). sfiA and sfiB (sfi for suppressor of filament induction) are identical to sulA and suiB, respectively, and were isolated as spontaneous thermoresistant revertants of tif-1 (recA441) Ion strains (15,23,24). The tif-l Ion strains filament and die at 41°C. To account for their data, George et al. (15) proposed that a division inhibitor (product of the sulA or sulB gene?) was induced by UV and that it might be more stable in Ion strains.By means of a Mu d(amp-lac) operon fusion that linked the structural gene for 3...

show abstract

“…A gene bank of C. trachomatis (serovar LI) DNA was prepared in the phage vector X1059. Choice of this vector was influenced by (i) the ease with which phage plaques, in contrast to bacterial colonies, can be screened by an in situ radioimmunoassay (26), (ii) the reported toxicity of outer membrane proteins for E. coli when present on plasmid vectors (10), and (iii) the previously reported expression of a chlamydial antigen in this system (27). From this library 20 clones exhibiting various degrees of seroreactivity with serum (IHS) from a patient with a C. trachomatis L1 infection were selected.…”

Section: Discussionmentioning

confidence: 99%

Molecular cloning of the major outer membrane protein of Chlamydia trachomatis

Allan¹,

Cunningham²,

Lovett³

1984

Infect Immun

View full text Add to dashboard Cite

A gene library of Chiamydia trachomatis (serovar LI) DNA has been prepared in the phage vector X1059.From this bank, 20 recombinant phage-expressing components which reacted with serum from a patient with a C. trachomatis (Ll) infection were chosen. Selective expression and radiolabeling of phage polypeptides in irradiated Escherichia coli demonstrated that one of these clones encoded a polypeptide doublet with an apparent molecular weight similar to that of the C. trachomatis (Ll) major outer membrane protein. Both species of this cloned doublet (40 and 41 kilodaltons) could be immunoprecipitated by serum from a patient with a C. trachomatis (LI) infection but not by normal human serum. Components of this apparent molecular weight were not precipitated from irradiated E. coli infected with vector phage X1059 by either of these sera. Comparison of the Staphylococcus aureus-V8 protease peptide maps of these two cloned polypeptides and chlamydial major outer membrane protein extracted from elementary bodies showed all three polypeptides to produce peptide fragments of 15.5, 13.8, and 11.5 kilodaltons. Due to the indentical apparent molecular weights of the fragments produced from the 40and 41-kilodalton cloned polypeptides, these were concluded to be different conformational forms of the same molecular species. These cloned components were indistinguishable from C. trachomatis (LI) ma,jor outer membrane protein.

show abstract

Cloned DNA fragment specifying major outer membrane protein a in Escherichia coli K-12

Cited by 35 publications

References 34 publications

Escherichia coli K-12 mutants that lack major outer membrane protein a

Escherichia coli K-12 mutants that lack major outer membrane protein a

Regulation of cell division in Escherichia coli: SOS induction and cellular location of the sulA protein, a key to lon-associated filamentation and death

Molecular cloning of the major outer membrane protein of Chlamydia trachomatis

Contact Info

Product

Resources

About