• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
J Bacteriol. Aug 1995; 177(15): 4481–4487.
PMCID: PMC177200

Identification and sequence analysis of genes involved in late steps in cobalamin (vitamin B12) synthesis in Rhodobacter capsulatus.


A 6.4-kb region of a 6.8-kb BamHI fragment carrying Rhodobacter capsulatus genes involved in late steps of cobalamin synthesis has been sequenced. The nucleotide sequence and genetic analysis revealed that this fragment contains eight genes arranged in at least three operons. Five of these eight genes show homology to genes involved in the cobalamin synthesis of Pseudomonas denitrificans and Salmonella typhimurium. The arrangement of these homologous genes differs considerably in the three genera. Upstream of five overlapping genes (named bluFEDCB), a promoter activity could be detected by using lacZ fusions. This promoter shows no regulation by oxygen, vitamin B12 (cobalamin), or cobinamide. Disruption of the bluE gene by a Tn5 insertion (strain AH2) results in reduced expression of the puf and puc operons, which encode pigment-binding proteins of the photosynthetic apparatus. The mutant strain AH2 can be corrected to a wild-type-like phenotype by addition of vitamin B12 or cobinamide dicyanide. Disruption of the bluB gene by an interposon (strain BB1) also disturbs the formation of the photosynthetic apparatus. The mutation of strain BB1 can be corrected by vitamin B12 but not by cobinamide. We propose that a lack of cobalamin results in deregulation and a decreased formation of the photosynthetic apparatus.

Full Text

The Full Text of this article is available as a PDF (413K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Andersson DI, Roth JR. Redox regulation of the genes for cobinamide biosynthesis in Salmonella typhimurium. J Bacteriol. 1989 Dec;171(12):6734–6739. [PMC free article] [PubMed]
  • Battersby AR. How nature builds the pigments of life: the conquest of vitamin B12. Science. 1994 Jun 10;264(5165):1551–1557. [PubMed]
  • Bauer CE, Bollivar DW, Suzuki JY. Genetic analyses of photopigment biosynthesis in eubacteria: a guiding light for algae and plants. J Bacteriol. 1993 Jul;175(13):3919–3925. [PMC free article] [PubMed]
  • Berg DE, Weiss A, Crossland L. Polarity of Tn5 insertion mutations in Escherichia coli. J Bacteriol. 1980 May;142(2):439–446. [PMC free article] [PubMed]
  • Brey RN, Banner CD, Wolf JB. Cloning of multiple genes involved with cobalamin (Vitamin B12) biosynthesis in Bacillus megaterium. J Bacteriol. 1986 Aug;167(2):623–630. [PMC free article] [PubMed]
  • Casadaban MJ, Cohen SN. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol. 1980 Apr;138(2):179–207. [PubMed]
  • Cauthen SE, Pattison JR, Lascelles J. Vitamin B(12) in photosynthetic bacteria and methionine synthesis by Rhodopseudomonas spheroides. Biochem J. 1967 Mar;102(3):774–781. [PMC free article] [PubMed]
  • Crouzet J, Cauchois L, Blanche F, Debussche L, Thibaut D, Rouyez MC, Rigault S, Mayaux JF, Cameron B. Nucleotide sequence of a Pseudomonas denitrificans 5.4-kilobase DNA fragment containing five cob genes and identification of structural genes encoding S-adenosyl-L-methionine: uroporphyrinogen III methyltransferase and cobyrinic acid a,c-diamide synthase. J Bacteriol. 1990 Oct;172(10):5968–5979. [PMC free article] [PubMed]
  • Crouzet J, Levy-Schil S, Cameron B, Cauchois L, Rigault S, Rouyez MC, Blanche F, Debussche L, Thibaut D. Nucleotide sequence and genetic analysis of a 13.1-kilobase-pair Pseudomonas denitrificans DNA fragment containing five cob genes and identification of structural genes encoding Cob(I)alamin adenosyltransferase, cobyric acid synthase, and bifunctional cobinamide kinase-cobinamide phosphate guanylyltransferase. J Bacteriol. 1991 Oct;173(19):6074–6087. [PMC free article] [PubMed]
  • Devereux J, Haeberli P, Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. [PMC free article] [PubMed]
  • Escalante-Semerena JC, Roth JR. Regulation of cobalamin biosynthetic operons in Salmonella typhimurium. J Bacteriol. 1987 May;169(5):2251–2258. [PMC free article] [PubMed]
  • Fellay R, Frey J, Krisch H. Interposon mutagenesis of soil and water bacteria: a family of DNA fragments designed for in vitro insertional mutagenesis of gram-negative bacteria. Gene. 1987;52(2-3):147–154. [PubMed]
  • Fonstein M, Haselkorn R. Chromosomal structure of Rhodobacter capsulatus strain SB1003: cosmid encyclopedia and high-resolution physical and genetic map. Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2522–2526. [PMC free article] [PubMed]
  • Aschauer HN, Fischer G, Isenberg KE, Meszaros K, Willinger U, Todd RD, Beran H, Strobl R, Lang M, Fuchs K, et al. No proof of linkage between schizophrenia-related disorders including schizophrenia and chromosome 2q21 region. Eur Arch Psychiatry Clin Neurosci. 1993;243(3-4):193–198. [PubMed]
  • Goldman BS, Roth JR. Genetic structure and regulation of the cysG gene in Salmonella typhimurium. J Bacteriol. 1993 Mar;175(5):1457–1466. [PMC free article] [PubMed]
  • Gorchein A. Magnesium protoporphyrin chelatase activity in Rhodopseudomonas spheroides. Studies with whole cells. Biochem J. 1972 Mar;127(1):97–106. [PMC free article] [PubMed]
  • Grabau C, Roth JR. A Salmonella typhimurium cobalamin-deficient mutant blocked in 1-amino-2-propanol synthesis. J Bacteriol. 1992 Apr;174(7):2138–2144. [PMC free article] [PubMed]
  • Gribskov M, Devereux J, Burgess RR. The codon preference plot: graphic analysis of protein coding sequences and prediction of gene expression. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):539–549. [PMC free article] [PubMed]
  • Hayashi Mitsunori, Kamikubo Tadashi. Isolation of 5,6-dimethylbenzimidazolyl cobamide coenzyme from Rhodopseudomonas spheroides. FEBS Lett. 1970 Oct 16;10(4):249–252. [PubMed]
  • Hübner P, Masepohl B, Klipp W, Bickle TA. nif gene expression studies in Rhodobacter capsulatus: ntrC-independent repression by high ammonium concentrations. Mol Microbiol. 1993 Oct;10(1):123–132. [PubMed]
  • Hübner P, Willison JC, Vignais PM, Bickle TA. Expression of regulatory nif genes in Rhodobacter capsulatus. J Bacteriol. 1991 May;173(9):2993–2999. [PMC free article] [PubMed]
  • Keen NT, Tamaki S, Kobayashi D, Trollinger D. Improved broad-host-range plasmids for DNA cloning in gram-negative bacteria. Gene. 1988 Oct 15;70(1):191–197. [PubMed]
  • Klug G. Regulation of expression of photosynthesis genes in anoxygenic photosynthetic bacteria. Arch Microbiol. 1993;159(5):397–404. [PubMed]
  • Ma D, Cook DN, O'Brien DA, Hearst JE. Analysis of the promoter and regulatory sequences of an oxygen-regulated bch operon in Rhodobacter capsulatus by site-directed mutagenesis. J Bacteriol. 1993 Apr;175(7):2037–2045. [PMC free article] [PubMed]
  • Marrs B. Genetic recombination in Rhodopseudomonas capsulata. Proc Natl Acad Sci U S A. 1974 Mar;71(3):971–973. [PMC free article] [PubMed]
  • Neidle EL, Kaplan S. 5-Aminolevulinic acid availability and control of spectral complex formation in hemA and hemT mutants of Rhodobacter sphaeroides. J Bacteriol. 1993 Apr;175(8):2304–2313. [PMC free article] [PubMed]
  • O'Toole GA, Trzebiatowski JR, Escalante-Semerena JC. The cobC gene of Salmonella typhimurium codes for a novel phosphatase involved in the assembly of the nucleotide loop of cobalamin. J Biol Chem. 1994 Oct 21;269(42):26503–26511. [PubMed]
  • Pollich M, Jock S, Klug G. Identification of a gene required for the oxygen-regulated formation of the photosynthetic apparatus of Rhodobacter capsulatus. Mol Microbiol. 1993 Nov;10(4):749–757. [PubMed]
  • Roth JR, Lawrence JG, Rubenfield M, Kieffer-Higgins S, Church GM. Characterization of the cobalamin (vitamin B12) biosynthetic genes of Salmonella typhimurium. J Bacteriol. 1993 Jun;175(11):3303–3316. [PMC free article] [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed]
  • von Gabain A, Belasco JG, Schottel JL, Chang AC, Cohen SN. Decay of mRNA in Escherichia coli: investigation of the fate of specific segments of transcripts. Proc Natl Acad Sci U S A. 1983 Feb;80(3):653–657. [PMC free article] [PubMed]
  • Yanisch-Perron C, Vieira J, Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. [PubMed]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...