Abstract
Two genes controlling motility functions in Bacillus subtilis were identified by DNA sequence analysis of a chromosomal fragment containing a strong promoter for sigma D RNA polymerase. Previous studies had shown that this sigma D-dependent promoter controls synthesis of a 1.6-kb transcript in vivo and in vitro. Sequence analysis revealed that the 1.6-kb transcript contains two open reading frames coding for protein sequences homologous to the Escherichia coli motA and motB gene products, respectively, and ends in a rho-independent termination site. Direct evidence linking these genes to motility functions in B. subtilis was obtained by precise localization by polymerase chain reaction of Tn917 transposon insertion mutations of Mot- strains, isolated by Zuberi et al. (A. R. Zuberi, C. Ying, H. M. Parker, and G. W. Ordal, J. Bacteriol. 172:6841-6848, 1990), to within this mot. operon. Replacement of each wild-type gene by in-frame deletion mutations yielded strains possessing paralyzed flagella and confirmed that both motA and motB are required for the motility of B. subtilis. These current findings support our earlier suggestions that sigma D in B. subtilis plays a central role in the control of gene expression for flagellar assembly, chemotaxis, and motility functions. Sigma F, the enteric homolog of sigma D, controls similar functions in E. coli and Salmonella typhimurium, and these factors appear to be representative of a family of factors implicated in flagellar synthesis in many bacterial species, which we propose to designate the sigma 28 family.
References
54
Referenced
55
-
Albertini , A. M. , T. Caramori , W. D. Crabb , F. Scoffone , and A. Galizzi . 1991 . TheflaA locus of Bacillus subtilis is part of a large operon coding for flagellar structures, motility functions, and an ATPase-like polypeptide . J. Bacteriol. 173 : 3573 - 3579 .
(
10.1128/JB.173.11.3573-3579.1991
) / J. Bacteriol. / TheflaA locus of Bacillus subtilis is part of a large operon coding for flagellar structures, motility functions, and an ATPase-like polypeptide by Albertini A. M. (1991) -
Anagnostopoulos , C. , and J. Spizizen . 1961 . Requirements for transformation in Bacillus subtilis . J. Bacteriol. 81 : 741 - 746 .
(
10.1128/JB.81.5.741-746.1961
) / J. Bacteriol. / Requirements for transformation in Bacillus subtilis by Anagnostopoulos C. (1961) -
Arnosti , D. N. , and M. J. Chamberlin . 1989 . Secondary v factor controls transcription of flagellar and chemotaxis genes in Eschenchia coli . Proc. Natl. Acad. Sci. USA 86 : 830 - 834 .
(
10.1073/pnas.86.3.830
) / Proc. Natl. Acad. Sci. USA / Secondary v factor controls transcription of flagellar and chemotaxis genes in Eschenchia coli by Arnosti D. N. (1989) - Ausubel F. M. R. Brent R. E. Kingston D. D. Moore J. A. Smith J. G. Seidman and K. Struhl. 1987. Current protocols in molecular biology. John Wiley & Sons Inc. New York.
-
Bartlett , D. H. , B. B. Frantz , and P. Matsumura . 1988 . Flagellar transcriptional activators FlbB and FlaI: gene sequences and 5' consensus sequences of operons under FlbB and FlaI control . J. Bacteriol. 170 : 1575 - 1581 .
(
10.1128/JB.170.4.1575-1581.1988
) / J. Bacteriol. / Flagellar transcriptional activators FlbB and FlaI: gene sequences and 5' consensus sequences of operons under FlbB and FlaI control by Bartlett D. H. (1988) -
Birnboim , H. C. , and J. Doly . 1979 . A rapid alkaline extraction procedure for screening recombinant plasmid DNA . Nucleic Acids Res. 7 : 1513 - 1523 .
(
10.1093/nar/7.6.1513
) / Nucleic Acids Res. / A rapid alkaline extraction procedure for screening recombinant plasmid DNA by Birnboim H. C. (1979) -
Brendel , V. , and E. N. Trifonov . 1984 . A computer algorithm for testing potential prokaryotic terminators . Nucleic Acids Res. 12 : 4411 - 4427 .
(
10.1093/nar/12.10.4411
) / Nucleic Acids Res. / A computer algorithm for testing potential prokaryotic terminators by Brendel V. (1984) -
Brutlag , D. L. , J. Clayton , P. Friedland , and L. H. Kedes . 1982 . SEQ: a nucleotide sequence analysis and recombination system . Nucleic Acids Res. 10 : 279 - 294 .
(
10.1093/nar/10.1.279
) / Nucleic Acids Res. / SEQ: a nucleotide sequence analysis and recombination system by Brutlag D. L. (1982) - Brutlag , D. L. , J. P. Dautricourt , S. Maulik , and J. Relph . 1990 . Improved sensitivity of biological sequence database searches . Comput. Appl. Biosci. 6 : 237 - 245 . / Comput. Appl. Biosci. / Improved sensitivity of biological sequence database searches by Brutlag D. L. (1990)
-
Chen , E. Y. , and P. H. Seeburg . 1985 . Laboratory methods. Supercoil sequencing: a fast and simple method for sequencing plasmid DNA . DNA 4 : 165 - 170 .
(
10.1089/dna.1985.4.165
) / DNA / Laboratory methods. Supercoil sequencing: a fast and simple method for sequencing plasmid DNA by Chen E. Y. (1985) -
Chun , S. Y. , and J. S. Parkinson . 1988 . Bacterial motility: membrane topology of the Escherichia coli MotB protein . Science 239 : 276 - 278 .
(
10.1126/science.2447650
) / Science / Bacterial motility: membrane topology of the Escherichia coli MotB protein by Chun S. Y. (1988) -
Connell , T. D. , W. J. Black , T. H. Kawula , D. S. Barritt , J. A. Dempsey , K. Kverneland , Jr. , A. Stephenson , B. S. Schepart , G. L. Murphy , and J. G. Cannon . 1988 . Recombination among protein II genes of Neisseria gonorrhoeae generates new coding sequences and increases structural variability in the protein II family . Mol. Microbiol. 2 : 227 - 236 .
(
10.1111/j.1365-2958.1988.tb00024.x
) / Mol. Microbiol. / Recombination among protein II genes of Neisseria gonorrhoeae generates new coding sequences and increases structural variability in the protein II family by Connell T. D. (1988) -
Dean , G. E. , R. M. Macnab , J. Stader , P. Matsumura , and C. Burks . 1984 . Gene sequence and predicted amino acid sequence of the motA protein, a membrane-associated protein required for flagellar rotation in Escherichia coli . J. Bacteriol. 159 : 991 - 999 .
(
10.1128/JB.159.3.991-999.1984
) / J. Bacteriol. / Gene sequence and predicted amino acid sequence of the motA protein, a membrane-associated protein required for flagellar rotation in Escherichia coli by Dean G. E. (1984) -
Ferrari , F. A. , E. Ferrari , and J. A. Hoch . 1982 . Chromosomal location of a Bacillus subtilis DNA fragment uniquely transcribed by sigma-28-containing RNA polymerase . J. Bacteriol. 152 : 780 - 785 .
(
10.1128/jb.152.2.780-785.1982
) / J. Bacteriol. / Chromosomal location of a Bacillus subtilis DNA fragment uniquely transcribed by sigma-28-containing RNA polymerase by Ferrari F. A. (1982) -
Gilman , M. Z. , and M. J. Chamberlin . 1983 . Developmental and genetic regulation of Bacillus subtilis genes transcribed by o-"-RNA polymerase . Cell 35 : 285 - 293 .
(
10.1016/0092-8674(83)90231-3
) / Cell / Developmental and genetic regulation of Bacillus subtilis genes transcribed by o-"-RNA polymerase by Gilman M. Z. (1983) -
Gilman , M. Z. , J. S. Glenn , V. L. Singer , and M. J. Chamberlin . 1984 . Isolation of sigma-28-specific promoters from Bacillus subtilis DNA . Gene 32 : 11 - 20 .
(
10.1016/0378-1119(84)90027-1
) / Gene / Isolation of sigma-28-specific promoters from Bacillus subtilis DNA by Gilman M. Z. (1984) -
Gilman , M. Z. , J. L. Wiggs , and M. J. Chamberlin . 1981 . Nucleotide sequences of two Bacillus subtilis promoters used by Bacillus subtilis sigma-28 RNA polymerase . Nucleic Acids Res. 9 : 5991 - 6000 .
(
10.1093/nar/9.22.5991
) / Nucleic Acids Res. / Nucleotide sequences of two Bacillus subtilis promoters used by Bacillus subtilis sigma-28 RNA polymerase by Gilman M. Z. (1981) -
Guerry , P. , S. M. Logan , S. Thornton , and T. J. Trust . 1990 . Genomic organization and expression of Campylobacter flagellin genes . J. Bacteriol. 172 : 1853 - 1860 .
(
10.1128/JB.172.4.1853-1860.1990
) / J. Bacteriol. / Genomic organization and expression of Campylobacter flagellin genes by Guerry P. (1990) -
Hager P. W. and J. C. Rabinowitz. 1985. Translational specificity in Bacillus subtilis p. 1-32. In D. A. Dubnau (ed.) The molecular biology of the bacilli vol. 2. Academic Press Inc. New York.
(
10.1016/B978-0-12-222702-8.50007-X
) -
Hanafusa , T. , A. Sakai , A. Tominaga , and M. Enomoto . 1989 . Isolation and characterization of Escherichia coli hag operator mutants whose hag 48 expression has become repressible by a Salmonella Hl repressor . Mol. Gen. Genet. 216 : 44 - 50 .
(
10.1007/BF00332229
) / Mol. Gen. Genet. / Isolation and characterization of Escherichia coli hag operator mutants whose hag 48 expression has become repressible by a Salmonella Hl repressor by Hanafusa T. (1989) -
Heidecker , G. , J. Messing , and B. Gronenborn . 1980 . A versatile primer for DNA sequencing in the M13mp 2 cloning system . Gene 10 : 69 - 73 .
(
10.1016/0378-1119(80)90145-6
) / Gene / A versatile primer for DNA sequencing in the M13mp 2 cloning system by Heidecker G. (1980) -
Helmann , J. D. 1991 . Alternative sigma factors and the regulation of flagellar gene expression . Mol. Microbiol. 5 : 2875 - 2882 .
(
10.1111/j.1365-2958.1991.tb01847.x
) / Mol. Microbiol. / Alternative sigma factors and the regulation of flagellar gene expression by Helmann J. D. (1991) -
Helmann , J. D. , and M. J. Chamberlin . 1987 . DNA sequence analysis suggests that expression of flagellar and chemotaxis genes in Eschenichia coli and Salmonella typhimurium is controlled by an alternative a factor . Proc. Natl. Acad. Sci. USA 84 : 6422 - 6424 .
(
10.1073/pnas.84.18.6422
) / Proc. Natl. Acad. Sci. USA / DNA sequence analysis suggests that expression of flagellar and chemotaxis genes in Eschenichia coli and Salmonella typhimurium is controlled by an alternative a factor by Helmann J. D. (1987) -
Helmann , J. D. , L. M. Marquez , and M. J. Chamberlin . 1988 . Cloning, sequencing, and disruption of the Bacillus subtilis e2 gene . J. Bacteriol. 170 : 1568 - 1574 .
(
10.1128/JB.170.4.1568-1574.1988
) / J. Bacteriol. / Cloning, sequencing, and disruption of the Bacillus subtilis e2 gene by Helmann J. D. (1988) -
Henikoff , S. 1987 . Unidirectional digestion with exonuclease III in DNA sequence analysis . Methods Enzymol. 155 : 156 - 165 .
(
10.1016/0076-6879(87)55014-5
) / Methods Enzymol. / Unidirectional digestion with exonuclease III in DNA sequence analysis by Henikoff S. (1987) - Innis M. A. D. H. Gelfand J. J. Sninsky and T. J. White. 1990. PCR protocols. Academic Press San Diego Calif.
-
Komeda , Y. 1986 . Transcriptional control of flagellar genes in Escherichia coli K-12 . J. Bacteriol. 168 : 1315 - 1318 .
(
10.1128/JB.168.3.1315-1318.1986
) / J. Bacteriol. / Transcriptional control of flagellar genes in Escherichia coli K-12 by Komeda Y. (1986) -
Kustu , S. , E. Santero , J. Keener , D. Popham , and D. Weiss . 1989 . Expression of ,4 (ntrA)-dependent genes is probably united by a common mechanism . Microbiol. Rev. 53 : 367 - 376 .
(
10.1128/mr.53.3.367-376.1989
) / Microbiol. Rev. / Expression of ,4 (ntrA)-dependent genes is probably united by a common mechanism by Kustu S. (1989) -
Kutsukake , K. , Y. Ohya , and T. Iino . 1990 . Transcriptional analysis of the flagellar regulon of Salmonella typhimurium . J. Bacteriol. 172 : 741 - 747 .
(
10.1128/JB.172.2.741-747.1990
) / J. Bacteriol. / Transcriptional analysis of the flagellar regulon of Salmonella typhimurium by Kutsukake K. (1990) -
Kyte , J. , and R. F. Doolittle . 1982 . A simple method for displaying the hydropathic character of a protein . J. Mol. Biol. 157 : 105 - 132 .
(
10.1016/0022-2836(82)90515-0
) / J. Mol. Biol. / A simple method for displaying the hydropathic character of a protein by Kyte J. (1982) - Macnab R. M. 1990. Genetics structure and assembly of the bacterial flagellum p. 77-106. In J. P. Armitage and J. M. Lackie (ed.) Biology of the chemotactic response vol. 46. Cambridge University Press Cambridge.
- Maniatis T. E. F. Fritsch and J. Sambrook. 1982. Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Cold Spring Harbor N.Y.
-
Marquez , L. M. , J. D. Helmann , E. Ferrari , H. M. Parker , G. W. Ordal , and M. J. Chamberlin . 1990 . Studies of edependent functions in Bacillus subtilis . J. Bacteriol. 172 : 3435 - 3443 .
(
10.1128/JB.172.6.3435-3443.1990
) / J. Bacteriol. / Studies of edependent functions in Bacillus subtilis by Marquez L. M. (1990) -
McLachlan , A. D. 1972 . Repeating sequences and gene duplication in proteins . J. Mol. Biol. 64 : 417 - 437 .
(
10.1016/0022-2836(72)90508-6
) / J. Mol. Biol. / Repeating sequences and gene duplication in proteins by McLachlan A. D. (1972) -
Mirel , D. B. , and M. J. Chamberlin . 1989 . The Bacillus subtilis flagellin gene (hag) is transcribed by the e8 form of RNA polymerase . J. Bacteriol. 171 : 3095 - 3101 .
(
10.1128/jb.171.6.3095-3101.1989
) / J. Bacteriol. / The Bacillus subtilis flagellin gene (hag) is transcribed by the e8 form of RNA polymerase by Mirel D. B. (1989) -
Nuiten , P. J. M. , F. J. A. M. van Asten , W. Gaastra , and B. A. M. van der Zeost . 1990 . Structural and functional analysis of two Campylobacter jejuni flagellin genes . J. Biol. Chem. 265 : 17798 - 17804 .
(
10.1016/S0021-9258(18)38234-6
) / J. Biol. Chem. / Structural and functional analysis of two Campylobacter jejuni flagellin genes by Nuiten P. J. M. (1990) -
Ohnishi , K. , K. Kutsukake , H. Suzuki , and T. lino. 1990 . Gene fliA encodes an alternative sigma factor specific for flagellar operons in Salmonella typhimurium . Mol. Gen. Genet. 221 : 139 - 147 .
(
10.1007/BF00261713
) / Mol. Gen. Genet. / Gene fliA encodes an alternative sigma factor specific for flagellar operons in Salmonella typhimurium by Ohnishi K. (1990) -
Perkins , J. B. , and P. J. Youngman . 1986 . Construction and properties of Tn 917-lac, a transposon derivative that mediates transcriptional gene fusions in Bacillus subtilis . Proc. Natl. Acad. Sci. USA 83 : 140 - 144 .
(
10.1073/pnas.83.1.140
) / Proc. Natl. Acad. Sci. USA / Construction and properties of Tn 917-lac, a transposon derivative that mediates transcriptional gene fusions in Bacillus subtilis by Perkins J. B. (1986) -
Pleier , E. , and R. Schmitt . 1989 . Identification and sequence analysis of two related flagellin genes in Rhizobium meliloti . J. Bacteriol. 171 : 1467 - 1475 .
(
10.1128/jb.171.3.1467-1475.1989
) / J. Bacteriol. / Identification and sequence analysis of two related flagellin genes in Rhizobium meliloti by Pleier E. (1989) -
Sanger , F. , S. Nicklen , and A. R. Coulson . 1977 . DNA sequencing with chain-terminating inhibitors . Proc. Natl. Acad. Sci. USA 74 : 5463 - 5467 .
(
10.1073/pnas.74.12.5463
) / Proc. Natl. Acad. Sci. USA / DNA sequencing with chain-terminating inhibitors by Sanger F. (1977) -
Silverman , M. , and M. Simon . 1976 . Operon controlling motility and chemotaxis in E. coli . Nature (London) 264 : 577 - 580 .
(
10.1038/264577a0
) / Nature (London) / Operon controlling motility and chemotaxis in E. coli by Silverman M. (1976) - Singer V. L. 1987. Ph.D. thesis. University of California Berkeley.
-
Southern , E. M. 1975 . Detection of specific sequences among DNA fragments separated by gel electrophoresis . J. Mol. Biol. 98 : 503 - 517 .
(
10.1016/S0022-2836(75)80083-0
) / J. Mol. Biol. / Detection of specific sequences among DNA fragments separated by gel electrophoresis by Southern E. M. (1975) -
Stader , J. , P. Matsumura , D. Vacante , G. E. Dean , and R. M. Macnab . 1986 . Nucleotide sequence of the Eschenichia coli motB gene and site-limited incorporation of its product into the cytoplasmic membrane . J. Bacteriol. 166 : 244 - 252 .
(
10.1128/JB.166.1.244-252.1986
) / J. Bacteriol. / Nucleotide sequence of the Eschenichia coli motB gene and site-limited incorporation of its product into the cytoplasmic membrane by Stader J. (1986) -
Stahl , M. L. , and E. Ferrari . 1984 . Replacement of the Bacillus subtilis subtilisin gene with an in vitro-derived deletion mutation . J. Bacteriol. 158 : 411 - 418 .
(
10.1128/jb.158.2.411-418.1984
) / J. Bacteriol. / Replacement of the Bacillus subtilis subtilisin gene with an in vitro-derived deletion mutation by Stahl M. L. (1984) -
Totten , P. A. , and S. Lory . 1990 . Characterization of the type a flagellin gene from Pseudomonas aeruginosa PAK . J. Bacteriol. 172 : 7188 - 7199 .
(
10.1128/JB.172.12.7188-7199.1990
) / J. Bacteriol. / Characterization of the type a flagellin gene from Pseudomonas aeruginosa PAK by Totten P. A. (1990) -
Wiggs , J. L. , M. Z. Gilman , and M. J. Chamberlin . 1981 . Heterogeneity of RNA polymerase in Bacillus subtilis: evidence for an additional cr factor in vegetative cells . Proc. Natl. Acad. Sci. USA 78 : 2762 - 2766 .
(
10.1073/pnas.78.5.2762
) / Proc. Natl. Acad. Sci. USA / Heterogeneity of RNA polymerase in Bacillus subtilis: evidence for an additional cr factor in vegetative cells by Wiggs J. L. (1981) -
Wilson , M. L. , and R. M. Macnab . 1988 . Overproduction of the MotA protein of Escherichia coli and estimation of its wild-type level . J. Bacteriol. 170 : 588 - 597 .
(
10.1128/JB.170.2.588-597.1988
) / J. Bacteriol. / Overproduction of the MotA protein of Escherichia coli and estimation of its wild-type level by Wilson M. L. (1988) -
Wilson , M. L. , and R. M. Macnab . 1990 . Co-overproduction and localization of the Escherichia coli motility proteins MotA and MotB . J. Bacteriol. 172 : 3932 - 3939 .
(
10.1128/JB.172.7.3932-3939.1990
) / J. Bacteriol. / Co-overproduction and localization of the Escherichia coli motility proteins MotA and MotB by Wilson M. L. (1990) - Yager T. D. and P. H. von Hippel. 1987. Transcript elongation and termination in Escherichia coli p. 1241-1275. In F. C. Neidhardt J. L. Ingraham K. B. Low B. Magasanik M. Schaechter and H. E. Umbarger (ed.) Escherichia coli and Salmonella typhimurium: cellular and molecular biology. American Society for Microbiology Washington D.C.
-
Yang , M. Y. , E. Ferrari , and D. J. Henner . 1984 . Cloning of the neutral protease gene of Bacillus subtilis and the use of the cloned gene to create an in vitro-derived deletion mutation . J. Bacteriol. 160 : 15 - 21 .
(
10.1128/JB.160.1.15-21.1984
) / J. Bacteriol. / Cloning of the neutral protease gene of Bacillus subtilis and the use of the cloned gene to create an in vitro-derived deletion mutation by Yang M. Y. (1984) -
Yanisch-Perron , C. , J. Vieira , and J. Messing . 1985 . Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mpl 8 and pUC19 vectors . Gene 33 : 103 - 119 .
(
10.1016/0378-1119(85)90120-9
) / Gene / Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mpl 8 and pUC19 vectors by Yanisch-Perron C. (1985) -
Zuberi , A. R. , C. Ying , D. S. Bischoff , and G. W. Ordal . 1991 . Gene-protein relationships in the flagellar hook-basal body complex of Bacillus subtilis: sequences of the flgB, flgC, flgG, fliE, and fliF genes . Gene 101 : 23 - 31 .
(
10.1016/0378-1119(91)90220-6
) / Gene / Gene-protein relationships in the flagellar hook-basal body complex of Bacillus subtilis: sequences of the flgB, flgC, flgG, fliE, and fliF genes by Zuberi A. R. (1991) -
Zuberi , A. R. , C. Ying , H. M. Parker , and G. W. Ordal . 1990 . Transposon Tn 9171acZ mutagenesis of Bacillus subtilis: identification of two new loci required for motility and chemotaxis . J. Bacteriol. 172 : 6841 - 6848 .
(
10.1128/JB.172.12.6841-6848.1990
) / J. Bacteriol. / Transposon Tn 9171acZ mutagenesis of Bacillus subtilis: identification of two new loci required for motility and chemotaxis by Zuberi A. R. (1990)
Dates
Type | When |
---|---|
Created | 8 years, 9 months ago (Nov. 7, 2016, 11:17 a.m.) |
Deposited | 3 years, 1 month ago (July 12, 2022, 1:15 p.m.) |
Indexed | 1 year, 4 months ago (April 7, 2024, 1:56 a.m.) |
Issued | 33 years, 1 month ago (July 1, 1992) |
Published | 33 years, 1 month ago (July 1, 1992) |
Published Print | 33 years, 1 month ago (July 1, 1992) |
@article{Mirel_1992, title={An operon of Bacillus subtilis motility genes transcribed by the sigma D form of RNA polymerase}, volume={174}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.174.13.4197-4204.1992}, DOI={10.1128/jb.174.13.4197-4204.1992}, number={13}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Mirel, D B and Lustre, V M and Chamberlin, M J}, year={1992}, month=jul, pages={4197–4204} }