Molecular characterization of the Salmonella typhimurium flhB operon and its protein products

T. Minamino, T. Iino, K. Kutsukake

Research output: Contribution to journalArticle

87 Citations (Scopus)

Abstract

The flhB and flhA genes constitute an operon called flhB operon on the Salmonella typhimurium chromosome. Their gene products are required for formation of the rod structure of flagellar apparatus. Furthermore, several lines of evidence suggest that they, together with FliI and FliH, may constitute the export apparatus of flagellin, the component protein of flagellar filament. In this study, we determined the nucleotide sequence of the entire flhB operon from S. typhimurium. It was shown that the flhB and flhA genes encode highly hydrophobic polypeptides with calculated molecular masses of 42,322 and 74,848 Da, respectively. Both proteins have several potential membrane-spanning segments, suggesting that they may be integral membrane proteins. The flhB operon was found to contain an additional open reading frame capable of encoding a polypeptide with a calculated molecular mass of 14,073 Da. We designated this open reading frame flhE. The N- terminal 16 amino acids of FlhE displays a feature of a typical signal sequence. A maxicell labeling experiment enabled us to identify the precursor and mature forms of the flhE gene products. Insertion of a kanamycin- resistant gene cartridge into the chromosomal flhE gene did not affect the motility of the cells, indicating that the flhE gene is not essential for flagellar formation and function. We have overproduced and purified N- terminally truncated FlhB and FlhA proteins and raised antibodies against them. By use of these antibodies, localization of the FlhB and FlhA proteins was analyzed by Western blotting (immunoblotting) with the fractionated cell extracts. The results obtained indicated that both proteins are localized in the cytoplasmic membrane.

Original languageEnglish
Pages (from-to)7630-7637
Number of pages8
JournalJournal of Bacteriology
Volume176
Issue number24
Publication statusPublished - 1994
Externally publishedYes

Fingerprint

Salmonella typhimurium
Operon
Genes
Proteins
Open Reading Frames
Western Blotting
Flagellin
Peptides
Kanamycin
Antibodies
Protein Sorting Signals
Cell Extracts
Membrane Potentials
Cell Movement
Membrane Proteins
Chromosomes
Cell Membrane
Amino Acids

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

Cite this

Molecular characterization of the Salmonella typhimurium flhB operon and its protein products. / Minamino, T.; Iino, T.; Kutsukake, K.

In: Journal of Bacteriology, Vol. 176, No. 24, 1994, p. 7630-7637.

Research output: Contribution to journalArticle

Minamino, T, Iino, T & Kutsukake, K 1994, 'Molecular characterization of the Salmonella typhimurium flhB operon and its protein products', Journal of Bacteriology, vol. 176, no. 24, pp. 7630-7637.
Minamino, T. ; Iino, T. ; Kutsukake, K. / Molecular characterization of the Salmonella typhimurium flhB operon and its protein products. In: Journal of Bacteriology. 1994 ; Vol. 176, No. 24. pp. 7630-7637.
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