Abstract:
:RasP is a predicted intramembrane metalloprotease of Bacillus subtilis that has been proposed to cleave the stress response anti-sigma factors RsiW and RsiV, the cell division protein FtsL, and remnant signal peptides within their transmembrane segments. To provide evidence for direct effects of RasP on putative substrates, we developed a heterologous coexpression system. Since expression of catalytically inactive RasP E21A inhibited expression of other membrane proteins in Escherichia coli, we added extra transmembrane segments to RasP E21A, which allowed accumulation of most other membrane proteins. A corresponding active version of RasP appeared to promiscuously cleave coexpressed membrane proteins, except those with a large periplasmic domain. However, stable cleavage products were not observed, even in clpP mutant E. coli Fusions of transmembrane segment-containing parts of FtsL and RsiW to E. coli maltose-binding protein (MBP) also resulted in proteins that appeared to be RasP substrates upon coexpression in E. coli, including FtsL with a full-length C-terminal domain (suggesting that prior cleavage by a site 1 protease is unnecessary) and RsiW designed to mimic the PrsW site 1 cleavage product (suggesting that further trimming by extracytoplasmic protease is unnecessary). Purified RasP cleaved His6-MBP-RsiW(73-118) in vitro within the RsiW transmembrane segment based on mass spectrometry analysis, demonstrating that RasP is an intramembrane protease. Surprisingly, purified RasP failed to cleave His6-MBP-FtsL(23-117). We propose that the lack of α-helix-breaking residues in the FtsL transmembrane segment creates a requirement for the membrane environment and/or an additional protein(s) in order for RasP to cleave FtsL.IMPORTANCE Intramembrane proteases govern important signaling pathways in nearly all organisms. In bacteria, they function in stress responses, cell division, pathogenesis, and other processes. Their membrane-associated substrates are typically inferred from genetic studies in the native bacterium. Evidence for direct effects has come sometimes from coexpression of the enzyme and potential substrate in a heterologous host and rarely from biochemical reconstitution of cleavage in vitro We applied these two approaches to the B. subtilis enzyme RasP and its proposed substrates RsiW and FtsL. We discovered potential pitfalls and solutions in heterologous coexpression experiments in E. coli, providing evidence that both substrates are cleaved by RasP in vivo but, surprisingly, that only RsiW was cleaved in vitro, suggesting that FtsL has an additional requirement.
journal_name
J Bacterioljournal_title
Journal of bacteriologyauthors
Parrell D,Zhang Y,Olenic S,Kroos Ldoi
10.1128/JB.00381-17subject
Has Abstractpub_date
2017-09-05 00:00:00issue
19eissn
0021-9193issn
1098-5530pii
JB.00381-17journal_volume
199pub_type
杂志文章abstract::The mechanisms by which Legionella pneumophila, a facultative intracellular parasite and the agent of Legionnaires' disease, acquires iron are largely unexplained. Several earlier studies indicated that L. pneumophila does not elaborate siderophores. However, we now present evidence that supernatants from L. pneumophi...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.182.3.749-757.2000
更新日期:2000-02-01 00:00:00
abstract::The Yersinia pestis proteome was studied as a function of temperature and calcium by two-dimensional differential gel electrophoresis. Over 4,100 individual protein spots were detected, of which hundreds were differentially expressed. A total of 43 differentially expressed protein spots, representing 24 unique protein...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.187.23.8172-8180.2005
更新日期:2005-12-01 00:00:00
abstract::Integrons are genetic elements that contribute to lateral gene transfer in bacteria as a consequence of possessing a site-specific recombination system. This system facilitates the spread of genes when they are part of mobile cassettes. Most integrons are contained within chromosomes and are confined to specific bacte...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.01950-05
更新日期:2006-08-01 00:00:00
abstract::SugE of Escherichia coli, first identified as a suppressor of groEL mutations but a member of the small multidrug resistance family, has not previously been shown to confer a drug resistance phenotype. We show that high-level expression of sugE leads to resistance to a subset of toxic quaternary ammonium compounds. ...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.184.9.2543-2545.2002
更新日期:2002-05-01 00:00:00
abstract::The gene (entD) encoding staphylococcal enterotoxin D (SED) has been located on a 27.6-kilobase penicillinase plasmid designated pIB485. This plasmid was present in all SED-producing strains tested. The entD gene was cloned on a 2.0-kilobase DNA fragment and was expressed in Escherichia coli. Sequence analysis of this...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.171.9.4799-4806.1989
更新日期:1989-09-01 00:00:00
abstract::A 40-kb region of DNA from Sorangium cellulosum So ce26, which contains polyketide synthase (PKS) genes for synthesis of the antifungal macrolide antibiotic soraphen A, was cloned. These genes were detected by homology to Streptomyces violaceoruber genes encoding components of granaticin PKS, thus extending this power...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.177.13.3673-3679.1995
更新日期:1995-07-01 00:00:00
abstract::Inhibition of sterol uptake in Saccharomyces cerevisiae sterol auxotroph FY3 (alpha hem1 erg7 ura) by delta-aminolevulinic acid (ALA) is dependent on the ability of the organism to synthesize heme from ALA. Sterol-depleted cells not exposed to ALA or strain PFY3 cells, with a double heme mutation, exposed to ALA did n...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.169.8.3707-3711.1987
更新日期:1987-08-01 00:00:00
abstract::Lysates of Escherichia coli Ymel obtained from cultures grown in the absence of tryptophan in minimal medium supplemented with 0.1% casein hydrolysate show an approximate fivefold increase in steady-state specific activity of both anthranilate synthetase and tryptophan synthetase A protein relative to cultures grown i...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.108.3.1181-1191.1971
更新日期:1971-12-01 00:00:00
abstract::Mutations in carbohydrate-negative mutants of Pseudomonas aeruginosa PAO1 individually deficient in glucose 6-phosphate dehydrogenase (zwf), 6-phosphogluconate dehydratase (edd), or pyruvate carboxylase (pyc) were mapped on the chromosome by plasmid R68.45-mediated conjugation and by bacteriophage F116L-mediated trans...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.156.3.1123-1129.1983
更新日期:1983-12-01 00:00:00
abstract::Sinorhizobium meliloti is a free-living soil bacterium which is capable of establishing a symbiotic relationship with the alfalfa plant (Medicago sativa). This symbiosis involves a network of bacterium-host signaling, as well as the potential for bacterium-bacterium communication, such as quorum sensing. In this study...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.184.13.3466-3475.2002
更新日期:2002-07-01 00:00:00
abstract::An alpha-L-arabinofuranosidase (EC 3.2.1.55) from the culture fluid of Rhodotorula flava IFO 0407 grown on beet arabinan as a carbon source has been highly purified. The purified enzyme has a pH optimum of 2.0. The enzyme is unusually acid stable, retaining 82% of its activity after being maintained for 24 h at pH 1.5...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.133.3.1073-1077.1978
更新日期:1978-03-01 00:00:00
abstract::Polysaccharide-containing material was located by histochemical staining on ultrathin sections of Bacillus cereus and several clostridial species during various phases of growth. These components were located along the cell wall in young vegetative cells and along the developing cortex in sporulating cells. In B. cere...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.98.3.1342-1354.1969
更新日期:1969-06-01 00:00:00
abstract::The major outer membrane lipoprotein (Lpp) of Escherichia coli possesses serine at position 2, which is thought to function as the outer membrane sorting signal, and lysine at the C terminus, through which Lpp covalently associates with peptidoglycan. Arginine (R) is present before the C-terminal lysine in the wild-ty...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.179.9.2857-2862.1997
更新日期:1997-05-01 00:00:00
abstract::Temperature-sensitive nalA mutants of Escherichia coli have been used to investigate the structure and functions of deoxyribonucleic acid (DNA) gyrase. Extracts of one such mutant (nalA43) had thermosensitive DNA gyrase subunit A activity but normal gyrase subunit B activity, proving definitively that nalA is the stru...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.140.2.424-435.1979
更新日期:1979-11-01 00:00:00
abstract::Lundgren, D. G. (Syracuse University, Syracuse, N.Y.) and J. J. Cooney. Chemical analyses of asporogenic mutants of Bacillus cereus. J. Bacteriol. 83:1287-1293. 1962.-Bacillus cereus ATCC 4342 and three temperature-sensitive asporogenic mutants were compared regarding some basic cellular components and response to met...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.83.6.1287-1293.1962
更新日期:1962-06-01 00:00:00
abstract::The gene encoding the outer membrane phosphate-selective porin protein P from Pseudomonas aeruginosa was cloned into Escherichia coli. The protein product was expressed and transported to the outer membrane of an E. coli phoE mutant and assembled into functional trimers. Expression of a product of the correct molecula...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.170.5.2312-2318.1988
更新日期:1988-05-01 00:00:00
abstract::The UspA1 and UspA2 proteins of Moraxella catarrhalis are structurally related, are exposed on the bacterial cell surface, and migrate as very high-molecular-weight complexes in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Previous analysis of uspA1 and uspA2 mutants of M. catarrhalis strain 035E indicat...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.182.5.1364-1373.2000
更新日期:2000-03-01 00:00:00
abstract::The nucleotide sequence of a 2.0-kilobase DNA segment containing the Clostridium acetobutylicum glnA gene was determined. The upstream region of the glnA gene contained two putative extended promoter consensus sequences (p1 and p2), characteristic of gram-positive bacteria. A third putative extended gram-positive prom...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.170.1.400-408.1988
更新日期:1988-01-01 00:00:00
abstract::Members of the carnobacteria have been extensively studied as probiotic cultures in aquacultures and protective cultures in seafood, diary, and meat. We report on the finished genome sequence of Carnobacterium sp. 17-4, which has been isolated from permanently cold seawater. The genetic information reveals a new circu...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.05113-11
更新日期:2011-07-01 00:00:00
abstract::ibeA is a virulence factor found in some extraintestinal pathogenic Escherichia coli (ExPEC) strains from the B2 phylogenetic group and particularly in newborn meningitic and avian pathogenic strains. It was shown to be involved in the invasion process of the newborn meningitic strain RS218. In a previous work, we sho...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.00089-12
更新日期:2012-09-01 00:00:00
abstract::Transcription of an aqpZ-lac fusion in a single copy on the Escherichia coli chromosome increased as cells entered the stationary growth phase. This was true in a variety of media, and increased transcription in enriched medium required the RpoS sigma factor. Expression of the aqpZ-lac fusion was not affected by up- o...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.184.15.4304-4307.2002
更新日期:2002-08-01 00:00:00
abstract::Purine nucleoside phosphorylase (EC 2.4.2.1) from Bacillus cereus T was examined at hourly intervals during growth and sporulation. The enzyme has maximal activity in extracts prepared from cells during stages I and II. The activity during exponential growth is only 6.6% of the maximum and that in free spores is only ...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.111.1.33-36.1972
更新日期:1972-07-01 00:00:00
abstract::Manganese is accumulated in Bacillus subtilis by a highly specific active transport system. This trace element "pump" is insensitive to added magnesium or calcium and preferentially accumulates manganese in the presence of cobalt, iron, and copper. Manganese uptake in B. subtilis is inhibited by cyanide, azide, pentac...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.113.3.1363-1372.1973
更新日期:1973-03-01 00:00:00
abstract::Plasmids for high-level expression of penicillin-binding protein 6 (PBP6) were constructed, giving rise to overproduction of PBP6 under the control of the lambda pR promoter in either the periplasmic or the cytoplasmic space. In contrast to penicillin-binding protein 5 (PBP5), the presence of high amounts of PBP6 in t...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.174.23.7572-7578.1992
更新日期:1992-12-01 00:00:00
abstract::Melanin production is a major virulence factor for Cryptococcus neoformans, an organism causing life-threatening infections in an estimated 10% of AIDS patients. In order to characterize the events involved in melanin synthesis, an enzyme having diphenol oxidase activity was purified and its gene was cloned. The enzym...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.176.3.656-664.1994
更新日期:1994-02-01 00:00:00
abstract::Mild alkaline extraction of native poly-beta-hydroxybutyrate (PHB) granules results in the solubilization of a protein fraction. Both the solubilized protein fraction and the extracted granules are essentially devoid of PHB synthetase activity unless recombined. The protein fraction has been separated by chromatograph...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.108.2.782-789.1971
更新日期:1971-11-01 00:00:00
abstract::Mutations in the tol-pal genes induce pleiotropic effects such as release of periplasmic proteins into the extracellular medium and hypersensitivity to drugs and detergents. Other outer membrane defective strains such as tolC, lpp, and rfa mutations are also altered in their outer membrane permeability. In this study,...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.180.18.4872-4878.1998
更新日期:1998-09-01 00:00:00
abstract::A RecA-independent chromosomal rearrangement in the upstream region of the streptolysin O (slo) gene of Streptococcus pyogenes which affects slo expression was identified. PCR analysis was used to demonstrate that this kind of rearrangement was found in several strains of different lineages. Chromosomal loci involved ...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.185.6.1857-1869.2003
更新日期:2003-03-01 00:00:00
abstract::Niederpruem, Donald J. (Indiana University Medical Center, Indianapolis). Respiration of basidiospores of Schizophyllum commune. J. Bacteriol. 88:210-215. 1964.-The aerobic metabolism of basidiospores of the wood-rotting mushroom Schizophyllum commune was investigated by use of manometric techniques and specific respi...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.88.1.210-215.1964
更新日期:1964-07-01 00:00:00
abstract::Escherichia coli K-12 strains contain a cytoplasmic activity, peptidase N, capable of hydrolyzing alanine-p-nitroanilide. Mutations in the structural gene for the enzyme, pepN, were mapped, and the properties of mutant strains were examined. The pepN locus lay between ompF and asnS at approximately 20.8 min on the E. ...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:
更新日期:1982-11-01 00:00:00