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. 1991 Nov;173(22):7113–7125. doi: 10.1128/jb.173.22.7113-7125.1991

Cloning and sequencing of a multigene family encoding the flagellins of Methanococcus voltae.

M L Kalmokoff 1, K F Jarrell 1
PMCID: PMC209217  PMID: 1718944

Abstract

The flagellins of Methanococcus voltae are encoded by a multigene family of four related genes (flaA, flaB1, flaB2, and flaB3). All four genes map within the same region of the genome, with the last three arranged in a direct tandem. Northern (RNA) blot and primer extension analyses of total cellular RNA indicate that all four genes are transcribed. The flaB1, flaB2, and flaB3 flagellins are transcribed as part of a large polycistronic message which encodes at least one more protein which is not a flagellin. An intercistronic stem-loop followed by a poly(T) tract located between the flaB2 and flaB3 genes appears to increase the resistance of the flaB1/flaB2 portion of this polycistronic message to digestion by endogenous RNases. The flaA gene, located approximately 600 bp upstream from the tandem, is transcribed as a separate message at very low levels. The four open reading frames encode proteins of molecular weights 23,900, 22,400, 22,800, and 25,500, much less than the Mr values of 33,000 and 31,000 for the flagellins calculated from sodium dodecyl sulfate-polyacrylamide gel electrophoresis of isolated flagellar filaments. Each flagellin contains multiple eukaryotic glycosylation signals (Arg-X-Ser/Thr), although they do not appear to be glycoproteins, and each has an 11- or 12-amino-acid leader peptide. The N termini of all four flagellins (amino acids 1 through 47 of the mature protein) are very hydrophobic, and this region shows a high degree of homology with the flagellins from Halobacterium halobium.

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Selected References

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  1. Arnosti D. N., Chamberlin M. J. Secondary sigma factor controls transcription of flagellar and chemotaxis genes in Escherichia coli. Proc Natl Acad Sci U S A. 1989 Feb;86(3):830–834. doi: 10.1073/pnas.86.3.830. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baier G., Piendl W., Redl B., Stöffler G. Structure, organization and evolution of the L1 equivalent ribosomal protein gene of the archaebacterium Methanococcus vannielii. Nucleic Acids Res. 1990 Feb 25;18(4):719–724. doi: 10.1093/nar/18.4.719. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Balch W. E., Fox G. E., Magrum L. J., Woese C. R., Wolfe R. S. Methanogens: reevaluation of a unique biological group. Microbiol Rev. 1979 Jun;43(2):260–296. doi: 10.1128/mr.43.2.260-296.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bollschweiler C., Kühn R., Klein A. Non-repetitive AT-rich sequences are found in intergenic regions of Methanococcus voltae DNA. EMBO J. 1985 Mar;4(3):805–809. doi: 10.1002/j.1460-2075.1985.tb03701.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brown J. W., Daniels C. J., Reeve J. N. Gene structure, organization, and expression in archaebacteria. Crit Rev Microbiol. 1989;16(4):287–338. doi: 10.3109/10408418909105479. [DOI] [PubMed] [Google Scholar]
  6. Chen C. Y., Beatty J. T., Cohen S. N., Belasco J. G. An intercistronic stem-loop structure functions as an mRNA decay terminator necessary but insufficient for puf mRNA stability. Cell. 1988 Feb 26;52(4):609–619. doi: 10.1016/0092-8674(88)90473-4. [DOI] [PubMed] [Google Scholar]
  7. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  8. Dharmavaram R., Gillevet P., Konisky J. Nucleotide sequence of the gene encoding the vanadate-sensitive membrane-associated ATPase of Methanococcus voltae. J Bacteriol. 1991 Mar;173(6):2131–2133. doi: 10.1128/jb.173.6.2131-2133.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Doetsch R. N., Sjoblad R. D. Flagellar structure and function in eubacteria. Annu Rev Microbiol. 1980;34:69–108. doi: 10.1146/annurev.mi.34.100180.000441. [DOI] [PubMed] [Google Scholar]
  10. Dunn R., McCoy J., Simsek M., Majumdar A., Chang S. H., Rajbhandary U. L., Khorana H. G. The bacteriorhodopsin gene. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6744–6748. doi: 10.1073/pnas.78.11.6744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gassmann G. S., Jacobs E., Deutzmann R., Göbel U. B. Analysis of the Borrelia burgdorferi GeHo fla gene and antigenic characterization of its gene product. J Bacteriol. 1991 Feb;173(4):1452–1459. doi: 10.1128/jb.173.4.1452-1459.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gerl L., Deutzmann R., Sumper M. Halobacterial flagellins are encoded by a multigene family. Identification of all five gene products. FEBS Lett. 1989 Feb 13;244(1):137–140. doi: 10.1016/0014-5793(89)81179-2. [DOI] [PubMed] [Google Scholar]
  13. Gerl L., Sumper M. Halobacterial flagellins are encoded by a multigene family. Characterization of five flagellin genes. J Biol Chem. 1988 Sep 15;263(26):13246–13251. [PubMed] [Google Scholar]
  14. Grogan D. W. Phenotypic characterization of the archaebacterial genus Sulfolobus: comparison of five wild-type strains. J Bacteriol. 1989 Dec;171(12):6710–6719. doi: 10.1128/jb.171.12.6710-6719.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Helmann J. D., Chamberlin M. J. DNA sequence analysis suggests that expression of flagellar and chemotaxis genes in Escherichia coli and Salmonella typhimurium is controlled by an alternative sigma factor. Proc Natl Acad Sci U S A. 1987 Sep;84(18):6422–6424. doi: 10.1073/pnas.84.18.6422. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Homma M., Fujita H., Yamaguchi S., Iino T. Regions of Salmonella typhimurium flagellin essential for its polymerization and excretion. J Bacteriol. 1987 Jan;169(1):291–296. doi: 10.1128/jb.169.1.291-296.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jarrell K. F., Koval S. F. Ultrastructure and biochemistry of Methanococcus voltae. Crit Rev Microbiol. 1989;17(1):53–87. doi: 10.3109/10408418909105722. [DOI] [PubMed] [Google Scholar]
  18. Jarrell K. F., Sprott G. D. The transmembrane electrical potential and intracellular pH in methanogenic bacteria. Can J Microbiol. 1981 Jul;27(7):720–728. doi: 10.1139/m81-110. [DOI] [PubMed] [Google Scholar]
  19. Joys T. M. The flagellar filament protein. Can J Microbiol. 1988 Apr;34(4):452–458. doi: 10.1139/m88-078. [DOI] [PubMed] [Google Scholar]
  20. Kalmokoff M. L., Jarrell K. F., Koval S. F. Isolation of flagella from the archaebacterium Methanococcus voltae by phase separation with Triton X-114. J Bacteriol. 1988 Apr;170(4):1752–1758. doi: 10.1128/jb.170.4.1752-1758.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kalmokoff M. L., Karnauchow T. M., Jarrell K. F. Conserved N-terminal sequences in the flagellins of archaebacteria. Biochem Biophys Res Commun. 1990 Feb 28;167(1):154–160. doi: 10.1016/0006-291x(90)91744-d. [DOI] [PubMed] [Google Scholar]
  22. Kostrzynska M., Betts J. D., Austin J. W., Trust T. J. Identification, characterization, and spatial localization of two flagellin species in Helicobacter pylori flagella. J Bacteriol. 1991 Feb;173(3):937–946. doi: 10.1128/jb.173.3.937-946.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kuwajima G., Asaka J., Fujiwara T., Fujiwara T., Node K., Kondo E. Nucleotide sequence of the hag gene encoding flagellin of Escherichia coli. J Bacteriol. 1986 Dec;168(3):1479–1483. doi: 10.1128/jb.168.3.1479-1483.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lechner J., Sumper M. The primary structure of a procaryotic glycoprotein. Cloning and sequencing of the cell surface glycoprotein gene of halobacteria. J Biol Chem. 1987 Jul 15;262(20):9724–9729. [PubMed] [Google Scholar]
  25. Macnab R. M., DeRosier D. J. Bacterial flagellar structure and function. Can J Microbiol. 1988 Apr;34(4):442–451. doi: 10.1139/m88-077. [DOI] [PubMed] [Google Scholar]
  26. Martin J. H., Savage D. C. Cloning, nucleotide sequence, and taxonomic implications of the flagellin gene of Roseburia cecicola. J Bacteriol. 1988 Jun;170(6):2612–2617. doi: 10.1128/jb.170.6.2612-2617.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. McKern N. M., O'Donnell I. J., Inglis A. S., Stewart D. J., Clark B. L. Amino acid sequence of pilin from Bacteroides nodosus (strain 198), the causative organism of ovine footrot. FEBS Lett. 1983 Nov 28;164(1):149–153. doi: 10.1016/0014-5793(83)80039-8. [DOI] [PubMed] [Google Scholar]
  28. Mullin D. A., Newton A. Ntr-like promoters and upstream regulatory sequence ftr are required for transcription of a developmentally regulated Caulobacter crescentus flagellar gene. J Bacteriol. 1989 Jun;171(6):3218–3227. doi: 10.1128/jb.171.6.3218-3227.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Murakawa G. J., Kwan C., Yamashita J., Nierlich D. P. Transcription and decay of the lac messenger: role of an intergenic terminator. J Bacteriol. 1991 Jan;173(1):28–36. doi: 10.1128/jb.173.1.28-36.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Pallesen L., Hindersson P. Cloning and sequencing of a Treponema pallidum gene encoding a 31.3-kilodalton endoflagellar subunit (FlaB2). Infect Immun. 1989 Jul;57(7):2166–2172. doi: 10.1128/iai.57.7.2166-2172.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Parales J., Jr, Greenberg E. P. N-terminal amino acid sequences and amino acid compositions of the Spirochaeta aurantia flagellar filament polypeptides. J Bacteriol. 1991 Feb;173(3):1357–1359. doi: 10.1128/jb.173.3.1357-1359.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Perlman D., Halvorson H. O. A putative signal peptidase recognition site and sequence in eukaryotic and prokaryotic signal peptides. J Mol Biol. 1983 Jun 25;167(2):391–409. doi: 10.1016/s0022-2836(83)80341-6. [DOI] [PubMed] [Google Scholar]
  33. Pleier E., Schmitt R. Identification and sequence analysis of two related flagellin genes in Rhizobium meliloti. J Bacteriol. 1989 Mar;171(3):1467–1475. doi: 10.1128/jb.171.3.1467-1475.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rosen K. M., Lamperti E. D., Villa-Komaroff L. Optimizing the northern blot procedure. Biotechniques. 1990 Apr;8(4):398–403. [PubMed] [Google Scholar]
  35. Sandman K., Krzycki J. A., Dobrinski B., Lurz R., Reeve J. N. HMf, a DNA-binding protein isolated from the hyperthermophilic archaeon Methanothermus fervidus, is most closely related to histones. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5788–5791. doi: 10.1073/pnas.87.15.5788. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sastry P. A., Pearlstone J. R., Smillie L. B., Paranchych W. Amino acid sequence of pilin isolated from pseudomonas aeruginosa PAK. FEBS Lett. 1983 Jan 24;151(2):253–256. doi: 10.1016/0014-5793(83)80080-5. [DOI] [PubMed] [Google Scholar]
  37. Shimmin L. C., Dennis P. P. Characterization of the L11, L1, L10 and L12 equivalent ribosomal protein gene cluster of the halophilic archaebacterium Halobacterium cutirubrum. EMBO J. 1989 Apr;8(4):1225–1235. doi: 10.1002/j.1460-2075.1989.tb03496.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Southam G., Kalmokoff M. L., Jarrell K. F., Koval S. F., Beveridge T. J. Isolation, characterization, and cellular insertion of the flagella from two strains of the archaebacterium Methanospirillum hungatei. J Bacteriol. 1990 Jun;172(6):3221–3228. doi: 10.1128/jb.172.6.3221-3228.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Sumper M., Berg E., Mengele R., Strobel I. Primary structure and glycosylation of the S-layer protein of Haloferax volcanii. J Bacteriol. 1990 Dec;172(12):7111–7118. doi: 10.1128/jb.172.12.7111-7118.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Thomashow L. S., Rittenberg S. C. Waveform analysis and structure of flagella and basal complexes from Bdellovibrio bacteriovorus 109J. J Bacteriol. 1985 Sep;163(3):1038–1046. doi: 10.1128/jb.163.3.1038-1046.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Totten P. A., Lara J. C., Lory S. The rpoN gene product of Pseudomonas aeruginosa is required for expression of diverse genes, including the flagellin gene. J Bacteriol. 1990 Jan;172(1):389–396. doi: 10.1128/jb.172.1.389-396.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Vogeli G., Kaytes P. S. Amplification, storage, and replication of libraries. Methods Enzymol. 1987;152:407–415. doi: 10.1016/0076-6879(87)52047-x. [DOI] [PubMed] [Google Scholar]
  43. Wallace R. B., Miyada C. G. Oligonucleotide probes for the screening of recombinant DNA libraries. Methods Enzymol. 1987;152:432–442. doi: 10.1016/0076-6879(87)52050-x. [DOI] [PubMed] [Google Scholar]
  44. Wieland F., Paul G., Sumper M. Halobacterial flagellins are sulfated glycoproteins. J Biol Chem. 1985 Dec 5;260(28):15180–15185. [PubMed] [Google Scholar]
  45. Woese C. R., Kandler O., Wheelis M. L. Towards a natural system of organisms: proposal for the domains Archaea, Bacteria, and Eucarya. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4576–4579. doi: 10.1073/pnas.87.12.4576. [DOI] [PMC free article] [PubMed] [Google Scholar]

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