Home LiteratureArticle Details
PMID: 9068642 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Identification and characterization of the ccdA gene, required for cytochrome c synthesis in Bacillus subtilis.

Journal of bacteriology ·Vol. 179 ·No. 6 ·1997-03-00 ·Pages 1962-73

Schiött T, von Wachenfeldt C, Hederstedt L

Abstract

The gram-positive, endospore-forming bacterium Bacillus subtilis contains several membrane-bound c-type cytochromes. We have isolated a mutant pleiotropically deficient in cytochromes c. The responsible mutation resides in a gene which we have named ccdA (cytochrome c defective). This gene is located at 173 degrees on the B. subtilis chromosome. The ccdA gene was found to be specifically required for synthesis of cytochromes of the c type. CcdA is a predicted 26-kDa integral membrane protein with no clear similarity to any known cytochrome c biogenesis protein but seems to be related to a part of Escherichia coli DipZ/DsbD. The ccdA gene is cotranscribed with two other genes. These genes encode a putative 13.5-kDa single-domain response regulator, similar to B. subtilis CheY and Spo0F, and a predicted 18-kDa hydrophobic protein with no similarity to any protein in databases, respectively. Inactivation of the three genes showed that only ccdA is required for cytochrome c synthesis. The results also demonstrated that cytochromes of the c type are not needed for growth of B. subtilis.

MeSH Terms
Amino Acid Sequence Bacillus subtilis/genetics,metabolism Bacterial Proteins Base Sequence Chromosome Mapping Cloning, Molecular Cytochrome c Group/biosynthesis Genes, Bacterial Membrane Proteins/chemistry,genetics,metabolism,physiology Molecular Sequence Data Mutation Open Reading Frames Protein Biosynthesis RNA, Bacterial/genetics RNA, Messenger/genetics Sequence Analysis, DNA
Chemicals
Bacterial Proteins CcdA protein, Bacillus Cytochrome c Group Membrane Proteins RNA, Bacterial RNA, Messenger
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Schiött T
Department of Microbiology, Lund University, Sweden.
von Wachenfeldt C
Hederstedt L
References (66)
66 references, click to expand
  1. Increased rate of asporogenous mutations following treatment of Bacillus subtilis spores with ethyl methanesulfonate.
    Mutat Res. 1971 Sep;13(1):93-6 PMID: 4999912
  2. Yeast TKL1 gene encodes a transketolase that is required for efficient glycolysis and biosynthesis of aromatic amino acids.
    J Biol Chem. 1993 Nov 15;268(32):24346-52 PMID: 8226984
  3. Alteration of haem-attachment and signal-cleavage sites for Paracoccus denitrificans cytochrome C550 probes pathway of c-type cytochrome biogenesis in Escherichia coli.
    Mol Microbiol. 1996 Mar;19(6):1193-204 PMID: 8730862
  4. The biogenesis of c-type cytochromes in Escherichia coli requires a membrane-bound protein, DipZ, with a protein disulphide isomerase-like domain.
    Mol Microbiol. 1995 Mar;15(6):1139-50 PMID: 7623667
  5. Escherichia coli K-12 restricts DNA containing 5-methylcytosine.
    Proc Natl Acad Sci U S A. 1986 Dec;83(23):9070-4 PMID: 3024165
  6. The cytochrome bc complex (menaquinone:cytochrome c reductase) in Bacillus subtilis has a nontraditional subunit organization.
    J Bacteriol. 1995 Dec;177(23):6751-60 PMID: 7592464
  7. Rapid and efficient cosmid cloning.
    Nucleic Acids Res. 1981 Jul 10;9(13):2989-98 PMID: 6269067
  8. The Bacillus subtilis hemAXCDBL gene cluster, which encodes enzymes of the biosynthetic pathway from glutamate to uroporphyrinogen III.
    J Bacteriol. 1991 Apr;173(8):2590-9 PMID: 1672867
  9. Identification and characterization of the tktB gene encoding a second transketolase in Escherichia coli K-12.
    J Bacteriol. 1993 Sep;175(17):5375-83 PMID: 8396116
  10. Fluorographic detection of radioactivity in polyacrylamide gels with the water-soluble fluor, sodium salicylate.
    Anal Biochem. 1979 Sep 15;98(1):132-5 PMID: 543547
  11. A mutation blocking the formation of membrane or periplasmic endogenous and exogenous c-type cytochromes in Escherichia coli permits the cytoplasmic formation of Hydrogenobacter thermophilus holo cytochrome c552.
    FEBS Lett. 1994 May 16;344(2-3):207-10 PMID: 8187885
  12. Sequence and characterization of Bacillus subtilis CheB, a homolog of Escherichia coli CheY, and its role in a different mechanism of chemotaxis.
    J Biol Chem. 1991 Jul 5;266(19):12301-5 PMID: 1905718
  13. New genes in the 170 degrees region of the Bacillus subtilis genome encode DNA gyrase subunits, a thioredoxin, a xylanase and an amino acid transporter.
    Microbiology. 1996 Nov;142 ( Pt 11):3097-101 PMID: 8969507
  14. An integrated map of the genome of the tubercle bacillus, Mycobacterium tuberculosis H37Rv, and comparison with Mycobacterium leprae.
    Proc Natl Acad Sci U S A. 1996 Apr 2;93(7):3132-7 PMID: 8610181
  15. Refined structure of transketolase from Saccharomyces cerevisiae at 2.0 A resolution.
    J Mol Biol. 1994 May 6;238(3):387-404 PMID: 8176731
  16. Nucleotide sequences of transcription and translation initiation regions in Bacillus phage phi 29 early genes.
    J Biol Chem. 1982 Jan 25;257(2):1053-62 PMID: 6274853
  17. Transformation in Bacillus subtilis. Fate of newly introduced transforming DNA.
    Mol Gen Genet. 1973;123(2):185-98 PMID: 4199220
  18. A survey of the Mycoplasma genitalium genome by using random sequencing.
    J Bacteriol. 1993 Dec;175(24):7918-30 PMID: 8253680
  19. The SWISS-PROT protein sequence data bank and its new supplement TREMBL.
    Nucleic Acids Res. 1996 Jan 1;24(1):21-5 PMID: 8594581
  20. Synthesis of holo Paracoccus denitrificans cytochrome c550 requires targeting to the periplasm whereas that of holo Hydrogenobacter thermophilus cytochrome c552 does not. Implications for c-type cytochrome biogenesis.
    FEBS Lett. 1994 Feb 28;340(1-2):65-70 PMID: 8119410
  21. Nucleotide sequence of the Escherichia coli K-12 transketolase (tkt) gene.
    Biochim Biophys Acta. 1993 Nov 16;1216(2):307-10 PMID: 8241274
  22. Regulation of aconitase synthesis in Bacillus subtilis: induction, feedback repression, and catabolite repression.
    J Bacteriol. 1974 Mar;117(3):1295-305 PMID: 4205196
  23. Revised assignment for the Bacillus subtilis spo0F gene and its homology with spo0A and with two Escherichia coli genes.
    Nucleic Acids Res. 1986 Jan 24;14(2):1063-72 PMID: 3003689
  24. Changes in the stability of specific mRNA species in response to growth stage in Bacillus subtilis.
    Proc Natl Acad Sci U S A. 1990 Nov;87(21):8355-9 PMID: 1700430
  25. Protein disulphide oxidoreductases in bacteria.
    Trends Biochem Sci. 1994 Apr;19(4):169-71 PMID: 8016867
  26. Cloning, nucleotide sequence, and expression of the Bacillus subtilis lon gene.
    J Bacteriol. 1994 Nov;176(21):6518-27 PMID: 7961402
  27. Identification and characterization of a new disulfide isomerase-like protein (DsbD) in Escherichia coli.
    EMBO J. 1995 Jul 17;14(14):3415-24 PMID: 7628442
  28. Molecular properties, genetics, and biosynthesis of Bacillus subtilis succinate dehydrogenase complex.
    Methods Enzymol. 1986;126:399-414 PMID: 3152413
  29. CotM of Bacillus subtilis, a member of the alpha-crystallin family of stress proteins, is induced during development and participates in spore outer coat formation.
    J Bacteriol. 1997 Mar;179(6):1887-97 PMID: 9068633
  30. Insertional mutagenesis in Bacillus subtilis: mechanism and use in gene cloning.
    Gene. 1982 Oct;19(3):277-84 PMID: 6295881
  31. Formation of several bacterial c-type cytochromes requires a novel membrane-anchored protein that faces the periplasm.
    Mol Microbiol. 1993 Aug;9(4):729-40 PMID: 8231805
  32. Bacillus subtilis 13-kilodalton cytochrome c-550 encoded by cccA consists of a membrane-anchor and a heme domain.
    J Biol Chem. 1990 Aug 15;265(23):13939-48 PMID: 2166045
  33. Whole-genome random sequencing and assembly of Haemophilus influenzae Rd.
    Science. 1995 Jul 28;269(5223):496-512 PMID: 7542800
  34. Discovery and sequence analysis of bacterial genes involved in the biogenesis of c-type cytochromes.
    J Biol Chem. 1991 Apr 25;266(12):7793-803 PMID: 1850420
  35. The cydD gene product, component of a heterodimeric ABC transporter, is required for assembly of periplasmic cytochrome c and of cytochrome bd in Escherichia coli.
    FEMS Microbiol Lett. 1994 Apr 1;117(2):217-23 PMID: 8181727
  36. Improved estimation of secondary structure in ribonucleic acids.
    Nat New Biol. 1973 Nov 14;246(150):40-1 PMID: 4519026
  37. Analysis of sporulation mutants. II. Mutants blocked in the citric acid cycle.
    J Bacteriol. 1968 Apr;95(4):1431-8 PMID: 4967197
  38. Role of His residues in Bacillus subtilis cytochrome b558 for haem binding and assembly of succinate: quinone oxidoreductase (complex II).
    Mol Microbiol. 1990 Jun;4(6):1045-56 PMID: 2120540
  39. Apo forms of cytochrome c550 and cytochrome cd1 are translocated to the periplasm of Paracoccus denitrificans in the absence of haem incorporation caused either mutation or inhibition of haem synthesis.
    Mol Microbiol. 1990 Jul;4(7):1181-92 PMID: 2172694
  40. Cytochrome c biogenesis in bacteria: a possible pathway begins to emerge.
    Mol Microbiol. 1994 Apr;12(1):1-9 PMID: 8057830
  41. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  42. The cycHJKL gene cluster plays an essential role in the biogenesis of c-type cytochromes in Bradyrhizobium japonicum.
    Mol Gen Genet. 1995 Apr 10;247(1):27-38 PMID: 7715601
  43. Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. strain PCC6803. I. Sequence features in the 1 Mb region from map positions 64% to 92% of the genome.
    DNA Res. 1995 Aug 31;2(4):153-66, 191-8 PMID: 8590279
  44. Escherichia coli genes required for cytochrome c maturation.
    J Bacteriol. 1995 Aug;177(15):4321-6 PMID: 7635817
  45. Basic local alignment search tool.
    J Mol Biol. 1990 Oct 5;215(3):403-10 PMID: 2231712
  46. Escherichia coli K-12 genes essential for the synthesis of c-type cytochromes and a third nitrate reductase located in the periplasm.
    Mol Microbiol. 1996 Feb;19(3):467-81 PMID: 8830238
  47. Cytochromes c biogenesis in a photosynthetic bacterium requires a periplasmic thioredoxin-like protein.
    Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2179-83 PMID: 8384715
  48. A comprehensive set of sequence analysis programs for the VAX.
    Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387-95 PMID: 6546423
  49. Bacillus subtilis citB gene is regulated synergistically by glucose and glutamine.
    J Bacteriol. 1985 Oct;164(1):155-64 PMID: 2413006
  50. Role of heme in synthesis and membrane binding of succinic dehydrogenase in Bacillus subtilis.
    J Bacteriol. 1979 May;138(2):377-82 PMID: 108259
  51. The effect of restriction on shotgun cloning and plasmid stability in Bacillus subtilis Marburg.
    Mol Gen Genet. 1987 Sep;209(2):335-42 PMID: 2823077
  52. The sigma factors of Bacillus subtilis.
    Microbiol Rev. 1995 Mar;59(1):1-30 PMID: 7708009
  53. TRANSFORMATION OF BIOCHEMICALLY DEFICIENT STRAINS OF BACILLUS SUBTILIS BY DEOXYRIBONUCLEATE.
    Proc Natl Acad Sci U S A. 1958 Oct 15;44(10):1072-8 PMID: 16590310
  54. An ordered collection of Bacillus subtilis DNA segments cloned in yeast artificial chromosomes.
    Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):6047-51 PMID: 8327482
  55. The minimal gene complement of Mycoplasma genitalium.
    Science. 1995 Oct 20;270(5235):397-403 PMID: 7569993
  56. Specific thiol compounds complement deficiency in c-type cytochrome biogenesis in Escherichia coli carrying a mutation in a membrane-bound disulphide isomerase-like protein.
    FEBS Lett. 1994 Oct 24;353(3):235-8 PMID: 7957865
  57. The functions and synthesis of bacterial c-type cytochromes with particular reference to Paracoccus denitrificans and Rhodobacter capsulatus.
    Biochim Biophys Acta. 1991 May 23;1058(1):17-20 PMID: 1646010
  58. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors.
    Gene. 1985;33(1):103-19 PMID: 2985470
  59. Molecular biology of Bacillus subtilis cytochromes.
    FEMS Microbiol Lett. 1992 Dec 15;100(1-3):91-100 PMID: 1335950
  60. Bacterial cytochromes c biogenesis.
    Genes Dev. 1992 Feb;6(2):268-83 PMID: 1310666
  61. Molecular weight determination of protein-dodecyl sulfate complexes by gel electrophoresis in a discontinuous buffer system.
    J Biol Chem. 1971 Oct 25;246(20):6328-34 PMID: 5127429
  62. Signal peptides: exquisitely designed transport promoters.
    Mol Microbiol. 1994 Sep;13(5):765-73 PMID: 7815936
  63. Isolation and characterization of an Escherichia coli mutant lacking cytochrome d terminal oxidase.
    J Bacteriol. 1983 Jun;154(3):1269-75 PMID: 6304009
  64. Bacillus subtilis cytochrome oxidase mutants: biochemical analysis and genetic evidence for two aa3-type oxidases.
    Mol Microbiol. 1991 Aug;5(8):2063-72 PMID: 1685007
  65. An essential role for DsbA in cytochrome c synthesis and formate-dependent nitrite reduction by Escherichia coli K-12.
    Arch Microbiol. 1995 Oct;164(4):301-7 PMID: 7487336
  66. Purification of aconitase from Bacillus subtilis and correlation of its N-terminal amino acid sequence with the sequence of the citB gene.
    J Bacteriol. 1987 Jul;169(7):3062-7 PMID: 3110133
Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.179.6.1962-1973.1997
Published
1997-03-00
Pages
1962-73
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC178920
Subset
IM
Databases
GENBANK
D64001, H10885, U00018, U32770, U32823, U38804, X77707, X87845, Z74024
External Links
PubMed source DOI: 10.1128/jb.179.6.1962-1973.1997 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2