Abstract
A 1.2-kb EcoRI genomic DNA fragment of Coxiella burnetti, when cloned onto a multicopy plasmid, was found to induce capsule synthesis (mucoidy) in Escherichia coli. Nucleotide sequence analysis revealed the presence of an open reading frame that could encode a protein of 270 amino acids. Insertion of a tet cassette into a unique NruI restriction site resulted in the loss of induction of mucoidy. Because of its ability to induce mucoidy, we designated this gene mucZ. Computer search for homologies to mucZ revealed 42% identity to an open reading frame located at 1 min of the E. coli chromosome. Interestingly, the C-terminal amino acid residues of MucZ share significant homology with the J domain of the DnaJ protein and its homologs, suggesting potential interactions between MucZ and components of the DnaK-chaperone machinery. Results presented in this paper suggest that E. coli requires DnaK-chaperone machinery for Lon-RcsA-mediated induction of capsule synthesis, as noticed first by S. Gottesman (personal communication). The induction caused by MucZ is independent of Lon-RcsA and is mediated through the two-component regulators RcsC and RcsB. DnaK and GrpE but not DnaJ are also required for the RcsB-mediated MucZ induction, and we propose that MucZ is a DnaJ-like chaperone protein that might be required for the formation of an active RcsA-RcsB complex and for the RcsC-dependent phosphorylation of RcsB. Discussions are presented that suggest three different roles for alternative forms of the DnaK-chaperone machinery in capsule production.
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- Bachmann B. J. Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev. 1972 Dec;36(4):525–557. doi: 10.1128/br.36.4.525-557.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Balbás P., Soberón X., Merino E., Zurita M., Lomeli H., Valle F., Flores N., Bolivar F. Plasmid vector pBR322 and its special-purpose derivatives--a review. Gene. 1986;50(1-3):3–40. doi: 10.1016/0378-1119(86)90307-0. [DOI] [PubMed] [Google Scholar]
- Bardwell J. C., Craig E. A. Major heat shock gene of Drosophila and the Escherichia coli heat-inducible dnaK gene are homologous. Proc Natl Acad Sci U S A. 1984 Feb;81(3):848–852. doi: 10.1073/pnas.81.3.848. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bardwell J. C., McGovern K., Beckwith J. Identification of a protein required for disulfide bond formation in vivo. Cell. 1991 Nov 1;67(3):581–589. doi: 10.1016/0092-8674(91)90532-4. [DOI] [PubMed] [Google Scholar]
- Bardwell J. C., Tilly K., Craig E., King J., Zylicz M., Georgopoulos C. The nucleotide sequence of the Escherichia coli K12 dnaJ+ gene. A gene that encodes a heat shock protein. J Biol Chem. 1986 Feb 5;261(4):1782–1785. [PubMed] [Google Scholar]
- Blum P., Ory J., Bauernfeind J., Krska J. Physiological consequences of DnaK and DnaJ overproduction in Escherichia coli. J Bacteriol. 1992 Nov;174(22):7436–7444. doi: 10.1128/jb.174.22.7436-7444.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brill J. A., Quinlan-Walshe C., Gottesman S. Fine-structure mapping and identification of two regulators of capsule synthesis in Escherichia coli K-12. J Bacteriol. 1988 Jun;170(6):2599–2611. doi: 10.1128/jb.170.6.2599-2611.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chen S. M., Takiff H. E., Barber A. M., Dubois G. C., Bardwell J. C., Court D. L. Expression and characterization of RNase III and Era proteins. Products of the rnc operon of Escherichia coli. J Biol Chem. 1990 Feb 15;265(5):2888–2895. [PubMed] [Google Scholar]
- Cherwonogrodzky J. W., Knodel M. H., Spence M. R. Increased encapsulation and virulence of Francisella tularensis live vaccine strain (LVS) by subculturing on synthetic medium. Vaccine. 1994 Jul;12(9):773–775. doi: 10.1016/0264-410x(94)90284-4. [DOI] [PubMed] [Google Scholar]
- Deretic V., Mohr C. D., Martin D. W. Mucoid Pseudomonas aeruginosa in cystic fibrosis: signal transduction and histone-like elements in the regulation of bacterial virulence. Mol Microbiol. 1991 Jul;5(7):1577–1583. doi: 10.1111/j.1365-2958.1991.tb01903.x. [DOI] [PubMed] [Google Scholar]
- Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Frosch M., Müller A. Phospholipid substitution of capsular polysaccharides and mechanisms of capsule formation in Neisseria meningitidis. Mol Microbiol. 1993 May;8(3):483–493. doi: 10.1111/j.1365-2958.1993.tb01592.x. [DOI] [PubMed] [Google Scholar]
- Gervais F. G., Drapeau G. R. Identification, cloning, and characterization of rcsF, a new regulator gene for exopolysaccharide synthesis that suppresses the division mutation ftsZ84 in Escherichia coli K-12. J Bacteriol. 1992 Dec;174(24):8016–8022. doi: 10.1128/jb.174.24.8016-8022.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gottesman S. Genetics of proteolysis in Escherichia coli*. Annu Rev Genet. 1989;23:163–198. doi: 10.1146/annurev.ge.23.120189.001115. [DOI] [PubMed] [Google Scholar]
- Gottesman S., Stout V. Regulation of capsular polysaccharide synthesis in Escherichia coli K12. Mol Microbiol. 1991 Jul;5(7):1599–1606. doi: 10.1111/j.1365-2958.1991.tb01906.x. [DOI] [PubMed] [Google Scholar]
- Gottesman S., Trisler P., Torres-Cabassa A. Regulation of capsular polysaccharide synthesis in Escherichia coli K-12: characterization of three regulatory genes. J Bacteriol. 1985 Jun;162(3):1111–1119. doi: 10.1128/jb.162.3.1111-1119.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hackstadt T., Williams J. C. Biochemical stratagem for obligate parasitism of eukaryotic cells by Coxiella burnetii. Proc Natl Acad Sci U S A. 1981 May;78(5):3240–3244. doi: 10.1073/pnas.78.5.3240. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hassett D. J., Woodruff W. A., Wozniak D. J., Vasil M. L., Cohen M. S., Ohman D. E. Cloning and characterization of the Pseudomonas aeruginosa sodA and sodB genes encoding manganese- and iron-cofactored superoxide dismutase: demonstration of increased manganese superoxide dismutase activity in alginate-producing bacteria. J Bacteriol. 1993 Dec;175(23):7658–7665. doi: 10.1128/jb.175.23.7658-7665.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kelly T., Dillard J. P., Yother J. Effect of genetic switching of capsular type on virulence of Streptococcus pneumoniae. Infect Immun. 1994 May;62(5):1813–1819. doi: 10.1128/iai.62.5.1813-1819.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Myers W. F., Baca O. G., Wisseman C. L., Jr Genome size of the rickettsia Coxiella burnetii. J Bacteriol. 1980 Oct;144(1):460–461. doi: 10.1128/jb.144.1.460-461.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Parker C. T., Kloser A. W., Schnaitman C. A., Stein M. A., Gottesman S., Gibson B. W. Role of the rfaG and rfaP genes in determining the lipopolysaccharide core structure and cell surface properties of Escherichia coli K-12. J Bacteriol. 1992 Apr;174(8):2525–2538. doi: 10.1128/jb.174.8.2525-2538.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
- Rubens C. E., Heggen L. M., Haft R. F., Wessels M. R. Identification of cpsD, a gene essential for type III capsule expression in group B streptococci. Mol Microbiol. 1993 May;8(5):843–855. doi: 10.1111/j.1365-2958.1993.tb01631.x. [DOI] [PubMed] [Google Scholar]
- Simpson J. A., Smith S. E., Dean R. T. Alginate inhibition of the uptake of Pseudomonas aeruginosa by macrophages. J Gen Microbiol. 1988 Jan;134(1):29–36. doi: 10.1099/00221287-134-1-29. [DOI] [PubMed] [Google Scholar]
- Sisk W. P., Chirikjian J. G., Lautenberger J., Jorcyk C., Papas T. S., Berman M. L., Zagursky R., Court D. L. A plasmid vector for cloning and expression of gene segments: expression of an HTLV-I envelope gene segment. Gene. 1986;48(2-3):183–193. doi: 10.1016/0378-1119(86)90076-4. [DOI] [PubMed] [Google Scholar]
- Sozhamannan S., Chattoraj D. K. Heat shock proteins DnaJ, DnaK, and GrpE stimulate P1 plasmid replication by promoting initiator binding to the origin. J Bacteriol. 1993 Jun;175(11):3546–3555. doi: 10.1128/jb.175.11.3546-3555.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Studier F. W. Genetic mapping of a mutation that causes ribonucleases III deficiency in Escherichia coli. J Bacteriol. 1975 Oct;124(1):307–316. doi: 10.1128/jb.124.1.307-316.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Takiff H. E., Chen S. M., Court D. L. Genetic analysis of the rnc operon of Escherichia coli. J Bacteriol. 1989 May;171(5):2581–2590. doi: 10.1128/jb.171.5.2581-2590.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ueguchi C., Kakeda M., Yamada H., Mizuno T. An analogue of the DnaJ molecular chaperone in Escherichia coli. Proc Natl Acad Sci U S A. 1994 Feb 1;91(3):1054–1058. doi: 10.1073/pnas.91.3.1054. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yura T., Mori H., Nagai H., Nagata T., Ishihama A., Fujita N., Isono K., Mizobuchi K., Nakata A. Systematic sequencing of the Escherichia coli genome: analysis of the 0-2.4 min region. Nucleic Acids Res. 1992 Jul 11;20(13):3305–3308. doi: 10.1093/nar/20.13.3305. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zuber M., Hoover T. A., Court D. L. Cloning, sequencing and expression of the dnaJ gene of Coxiella burnetii. Gene. 1995 Jan 11;152(1):99–102. doi: 10.1016/0378-1119(94)00687-n. [DOI] [PubMed] [Google Scholar]
- Zuber M., Hoover T. A., Powell B. S., Court D. L. Analysis of the rnc locus of Coxiella burnetii. Mol Microbiol. 1994 Oct;14(2):291–300. doi: 10.1111/j.1365-2958.1994.tb01290.x. [DOI] [PubMed] [Google Scholar]