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PMID: 2828336 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Characterization of the genes for the hexagonally arranged surface layer proteins in protein-producing Bacillus brevis 47: complete nucleotide sequence of the middle wall protein gene.

Journal of bacteriology ·Vol. 170 ·No. 2 ·1988-02-00 ·Pages 935-45

Tsuboi A, Uchihi R, Adachi T, Sasaki T, Hayakawa S, Yamagata H, Tsukagoshi N, Udaka S

Abstract

Bacillus brevis 47 contains two surface (S)-layer proteins, termed the outer wall protein (OWP) and the middle wall protein (MWP), which form a hexagonal array in the cell wall. The MWP and OWP genes are contained in the 9-kilobase-pair (kbp) BclI fragment and constitute an operon under coordinate control of their expression. The nucleotide sequence of a 3.8-kbp EcoRI-SacI fragment containing the entire MWP gene has been determined in this study. Together with the DNA sequence of the promoter region for the MWP-OWP gene operon (H. Yamagata, T. Adachi, A. Tsuboi, M. Takao, T. Sasaki, N. Tsukagoshi, and S. Udaka, J. Bacteriol. 169:1239-1245, 1987) and that of the OWP gene (A. Tsuboi, R. Uchihi, R. Tabata, Y. Takahashi, H. Hashiba, T. Sasaki, H. Yamagata, N. Tsukagoshi, and S. Udaka, J. Bacteriol. 168:365-373, 1986), the complete nucleotide sequence of the MWP-OWP gene operon has been determined. The MWP gene encodes a secretory precursor of the MWP, consisting of a total of 1,053 amino acid residues with a signal peptide of 23 amino acid residues at its amino-terminal end. Bacillus subtilis harboring the MWP gene synthesized an immunoreactive polypeptide with almost the same molecular weight as the authentic MWP, as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The amino acid compositions deduced from the MWP and OWP genes were similar to the chemical amino acid compositions of other S-layer proteins in the predominance of acidic amino acids compared with basic amino acids and in the very low content of sulfur-containing amino acids. The acidic nature of the MWP and OWP was confirmed by isoelectric focusing on polyacrylamide gels. In addition, circular dichroism spectra indicated that the S-layer proteins in B. brevis 47 were composed of approximately 30% beta-sheet and 5% alpha-helical structures, with the remainder of the polypeptide backbone being aperiodic in nature.

MeSH Terms
Amino Acid Sequence Bacillus/genetics,ultrastructure Bacterial Proteins/analysis,genetics Base Sequence Cell Wall/ultrastructure Cloning, Molecular Codon/genetics DNA Restriction Enzymes DNA, Bacterial/genetics Electrophoresis, Polyacrylamide Gel Gene Expression Regulation Genes, Bacterial Isoelectric Focusing Membrane Proteins/analysis,genetics Molecular Sequence Data Nucleic Acid Hybridization Operon Promoter Regions, Genetic
Chemicals
Bacterial Proteins Codon DNA, Bacterial Membrane Proteins DNA Restriction Enzymes
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Tsuboi A
Department of Food Science and Technology, Faculty of Agriculture, Nagoya University, Japan.
Uchihi R
Adachi T
Sasaki T
Hayakawa S
Yamagata H
Tsukagoshi N
Udaka S
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.170.2.935-945.1988
Published
1988-02-00
Pages
935-45
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC210745
Subset
IM
Databases
GENBANK
M19115
External Links
PubMed source DOI: 10.1128/jb.170.2.935-945.1988 PMC Full Text View journal details
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