Abstract
Insertion mutagenesis has been used to isolate Salmonella typhimurium strains that are blocked in the conversion of 5-aminolevulinic acid (ALA) to heme. These mutants define the steps of the heme biosynthetic pathway after ALA. Insertions were recovered at five unlinked loci: hemB, hemCD, and hemE, which have been mapped previously in S. typhimurium, and hemG and hemH, which have been described only for Escherichia coli. No other simple hem mutants were found. However, double mutants are described that are auxotrophic for heme during aerobic growth and fail to convert coproporphyrinogen III to protoporphyrinogen IX. These mutant strains are defective in two genes, hemN and hemF. Single mutants defective only in hemN require heme for anaerobic growth on glycerol plus nitrate but not for aerobic growth on glycerol. Mutants defective only in hemF have no apparent growth defect. We suggest that these two genes encode alternative forms of coproporphyrinogen oxidase. Anaerobic heme synthesis requires hemN function, while either hemN or hemF is sufficient for aerobic heme synthesis. These phenotypes are consistent with the requirement of a well-characterized class of coproporphyrinogen oxidase for molecular oxygen.
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Selected References
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- Adhya S., Cleary P., Campbell A. A deletion analysis of prophage lambda and adjacent genetic regions. Proc Natl Acad Sci U S A. 1968 Nov;61(3):956–962. doi: 10.1073/pnas.61.3.956. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Alper M. D., Ames B. N. Positive selection of mutants with deletions of the gal-chl region of the Salmonella chromosome as a screening procedure for mutagens that cause deletions. J Bacteriol. 1975 Jan;121(1):259–266. doi: 10.1128/jb.121.1.259-266.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Avissar Y. J., Beale S. I. Identification of the enzymatic basis for delta-aminolevulinic acid auxotrophy in a hemA mutant of Escherichia coli. J Bacteriol. 1989 Jun;171(6):2919–2924. doi: 10.1128/jb.171.6.2919-2924.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Avissar Y. J., Ormerod J. G., Beale S. I. Distribution of delta-aminolevulinic acid biosynthetic pathways among phototrophic bacterial groups. Arch Microbiol. 1989;151(6):513–519. doi: 10.1007/BF00454867. [DOI] [PubMed] [Google Scholar]
- BOGORAD L. The enzymatic synthesis of porphyrins from porphobilinogen. I. Uroporphyrin I. J Biol Chem. 1958 Aug;233(2):501–509. [PubMed] [Google Scholar]
- Berkowitz D., Hushon J. M., Whitfield H. J., Jr, Roth J., Ames B. N. Procedure for identifying nonsense mutations. J Bacteriol. 1968 Jul;96(1):215–220. doi: 10.1128/jb.96.1.215-220.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Breton R., Sanfaçon H., Papayannopoulos I., Biemann K., Lapointe J. Glutamyl-tRNA synthetase of Escherichia coli. Isolation and primary structure of the gltX gene and homology with other aminoacyl-tRNA synthetases. J Biol Chem. 1986 Aug 15;261(23):10610–10617. [PubMed] [Google Scholar]
- Castilho B. A., Olfson P., Casadaban M. J. Plasmid insertion mutagenesis and lac gene fusion with mini-mu bacteriophage transposons. J Bacteriol. 1984 May;158(2):488–495. doi: 10.1128/jb.158.2.488-495.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cauthen S. E., Foster M. A., Woods D. D. Methionine synthesis by extracts of Salmonella typhimurium. Biochem J. 1966 Feb;98(2):630–635. doi: 10.1042/bj0980630. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chumley F. G., Menzel R., Roth J. R. Hfr formation directed by tn10. Genetics. 1979 Apr;91(4):639–655. doi: 10.1093/genetics/91.4.639. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Claiborne A., Fridovich I. Purification of the o-dianisidine peroxidase from Escherichia coli B. Physicochemical characterization and analysis of its dual catalatic and peroxidatic activities. J Biol Chem. 1979 May 25;254(10):4245–4252. [PubMed] [Google Scholar]
- Claiborne A., Malinowski D. P., Fridovich I. Purification and characterization of hydroperoxidase II of Escherichia coli B. J Biol Chem. 1979 Nov 25;254(22):11664–11668. [PubMed] [Google Scholar]
- Cox R., Charles H. P. Porphyrin-accumulating mutants of Escherichia coli. J Bacteriol. 1973 Jan;113(1):122–132. doi: 10.1128/jb.113.1.122-132.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Demerec M., Adelberg E. A., Clark A. J., Hartman P. E. A proposal for a uniform nomenclature in bacterial genetics. Genetics. 1966 Jul;54(1):61–76. doi: 10.1093/genetics/54.1.61. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Desrochers M., Peloquin L., Săsărman A. Mapping of the hemE locus in Salmonella typhimurium. J Bacteriol. 1978 Sep;135(3):1151–1153. doi: 10.1128/jb.135.3.1151-1153.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Drolet M., Péloquin L., Echelard Y., Cousineau L., Sasarman A. Isolation and nucleotide sequence of the hemA gene of Escherichia coli K12. Mol Gen Genet. 1989 Apr;216(2-3):347–352. doi: 10.1007/BF00334375. [DOI] [PubMed] [Google Scholar]
- Echelard Y., Dymetryszyn J., Drolet M., Sasarman A. Nucleotide sequence of the hemB gene of Escherichia coli K12. Mol Gen Genet. 1988 Nov;214(3):503–508. doi: 10.1007/BF00330487. [DOI] [PubMed] [Google Scholar]
- Elliott T., Avissar Y. J., Rhie G. E., Beale S. I. Cloning and sequence of the Salmonella typhimurium hemL gene and identification of the missing enzyme in hemL mutants as glutamate-1-semialdehyde aminotransferase. J Bacteriol. 1990 Dec;172(12):7071–7084. doi: 10.1128/jb.172.12.7071-7084.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Elliott T. Cloning, genetic characterization, and nucleotide sequence of the hemA-prfA operon of Salmonella typhimurium. J Bacteriol. 1989 Jul;171(7):3948–3960. doi: 10.1128/jb.171.7.3948-3960.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Elliott T., Roth J. R. Characterization of Tn10d-Cam: a transposition-defective Tn10 specifying chloramphenicol resistance. Mol Gen Genet. 1988 Aug;213(2-3):332–338. doi: 10.1007/BF00339599. [DOI] [PubMed] [Google Scholar]
- Elliott T., Roth J. R. Heme-deficient mutants of Salmonella typhimurium: two genes required for ALA synthesis. Mol Gen Genet. 1989 Apr;216(2-3):303–314. doi: 10.1007/BF00334369. [DOI] [PubMed] [Google Scholar]
- Escalante-Semerena J. C., Roth J. R. Regulation of cobalamin biosynthetic operons in Salmonella typhimurium. J Bacteriol. 1987 May;169(5):2251–2258. doi: 10.1128/jb.169.5.2251-2258.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Frey B., McCloskey J., Kersten W., Kersten H. New function of vitamin B12: cobamide-dependent reduction of epoxyqueuosine to queuosine in tRNAs of Escherichia coli and Salmonella typhimurium. J Bacteriol. 1988 May;170(5):2078–2082. doi: 10.1128/jb.170.5.2078-2082.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grimm B., Bull A., Breu V. Structural genes of glutamate 1-semialdehyde aminotransferase for porphyrin synthesis in a cyanobacterium and Escherichia coli. Mol Gen Genet. 1991 Jan;225(1):1–10. doi: 10.1007/BF00282635. [DOI] [PubMed] [Google Scholar]
- Hughes K. T., Roth J. R. Transitory cis complementation: a method for providing transposition functions to defective transposons. Genetics. 1988 May;119(1):9–12. doi: 10.1093/genetics/119.1.9. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ingledew W. J., Poole R. K. The respiratory chains of Escherichia coli. Microbiol Rev. 1984 Sep;48(3):222–271. doi: 10.1128/mr.48.3.222-271.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Janzer J. J., Stan-Lotter H., Sanderson K. E. Isolation and characterization of hemin-permeable, envelope-defective mutants of Salmonella typhimurium. Can J Microbiol. 1981 Feb;27(2):226–237. doi: 10.1139/m81-034. [DOI] [PubMed] [Google Scholar]
- Jeter R. M., Olivera B. M., Roth J. R. Salmonella typhimurium synthesizes cobalamin (vitamin B12) de novo under anaerobic growth conditions. J Bacteriol. 1984 Jul;159(1):206–213. doi: 10.1128/jb.159.1.206-213.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jordan P. M., Mgbeje B. I., Alwan A. F., Thomas S. D. Nucleotide sequence of hemD, the second gene in the hem operon of Escherichia coli K-12. Nucleic Acids Res. 1987 Dec 23;15(24):10583–10583. doi: 10.1093/nar/15.24.10583. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jordan P. M., Mgbeje B. I., Thomas S. D., Alwan A. F. Nucleotide sequence for the hemD gene of Escherichia coli encoding uroporphyrinogen III synthase and initial evidence for a hem operon. Biochem J. 1988 Jan 15;249(2):613–616. doi: 10.1042/bj2490613. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Li J. M., Brathwaite O., Cosloy S. D., Russell C. S. 5-Aminolevulinic acid synthesis in Escherichia coli. J Bacteriol. 1989 May;171(5):2547–2552. doi: 10.1128/jb.171.5.2547-2552.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Li J. M., Russell C. S., Cosloy S. D. Cloning and structure of the hem A gene of Escherichia coli K-12. Gene. 1989 Oct 30;82(2):209–217. doi: 10.1016/0378-1119(89)90046-2. [DOI] [PubMed] [Google Scholar]
- Li J. M., Russell C. S., Cosloy S. D. The structure of the Escherichia coli hemB gene. Gene. 1989 Jan 30;75(1):177–184. doi: 10.1016/0378-1119(89)90394-6. [DOI] [PubMed] [Google Scholar]
- McConville M. L., Charles H. P. Isolation of haemin-requiring mutants of Escherichia coli K12. J Gen Microbiol. 1979 Jul;113(1):155–164. doi: 10.1099/00221287-113-1-155. [DOI] [PubMed] [Google Scholar]
- McConville M. L., Charles H. P. Mutants of Escherichia coli K12 permeable to haemin. J Gen Microbiol. 1979 Jul;113(1):165–168. doi: 10.1099/00221287-113-1-165. [DOI] [PubMed] [Google Scholar]
- Miyamoto K., Nakahigashi K., Nishimura K., Inokuchi H. Isolation and characterization of visible light-sensitive mutants of Escherichia coli K12. J Mol Biol. 1991 Jun 5;219(3):393–398. doi: 10.1016/0022-2836(91)90180-e. [DOI] [PubMed] [Google Scholar]
- O'Neill G. P., Chen M. W., Söll D. delta-Aminolevulinic acid biosynthesis in Escherichia coli and Bacillus subtilis involves formation of glutamyl-tRNA. FEMS Microbiol Lett. 1989 Aug;51(3):255–259. doi: 10.1016/0378-1097(89)90406-0. [DOI] [PubMed] [Google Scholar]
- Peters A. C., Wimpenny J. W., Coombs J. P. Oxygen profiles in, and in the agar beneath, colonies of Bacillus cereus, Staphylococcus albus and Escherichia coli. J Gen Microbiol. 1987 May;133(5):1257–1263. doi: 10.1099/00221287-133-5-1257. [DOI] [PubMed] [Google Scholar]
- Porra R. J., Falk J. E. The enzymic conversion of coproporphyrinogen 3 into protoporphyrin 9. Biochem J. 1964 Jan;90(1):69–75. doi: 10.1042/bj0900069. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Powell K. A., Cox R., McConville M., Charles H. P. Mutations affecting porphyrin biosynthesis in Escherichia coli. Enzyme. 1973;16(1):65–73. doi: 10.1159/000459363. [DOI] [PubMed] [Google Scholar]
- Roof D. M., Roth J. R. Ethanolamine utilization in Salmonella typhimurium. J Bacteriol. 1988 Sep;170(9):3855–3863. doi: 10.1128/jb.170.9.3855-3863.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SANO S., GRANICK S. Mitochondrial coproporphyrinogen oxidase and protoporphyrin formation. J Biol Chem. 1961 Apr;236:1173–1180. [PubMed] [Google Scholar]
- Sanderson K. E., Roth J. R. Linkage map of Salmonella typhimurium, edition VII. Microbiol Rev. 1988 Dec;52(4):485–532. doi: 10.1128/mr.52.4.485-532.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sasarman A., Nepveu A., Echelard Y., Dymetryszyn J., Drolet M., Goyer C. Molecular cloning and sequencing of the hemD gene of Escherichia coli K-12 and preliminary data on the Uro operon. J Bacteriol. 1987 Sep;169(9):4257–4262. doi: 10.1128/jb.169.9.4257-4262.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schmieger H. Phage P22-mutants with increased or decreased transduction abilities. Mol Gen Genet. 1972;119(1):75–88. doi: 10.1007/BF00270447. [DOI] [PubMed] [Google Scholar]
- Seehra J. S., Jordan P. M., Akhtar M. Anaerobic and aerobic coproporphyrinogen III oxidases of Rhodopseudomonas spheroides. Mechanism and stereochemistry of vinyl group formation. Biochem J. 1983 Mar 1;209(3):709–718. doi: 10.1042/bj2090709. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Straka J. G. High-performance liquid chromatography of porphyrin methyl esters. Methods Enzymol. 1986;123:352–363. doi: 10.1016/s0076-6879(86)23042-6. [DOI] [PubMed] [Google Scholar]
- Strauch K. L., Lenk J. B., Gamble B. L., Miller C. G. Oxygen regulation in Salmonella typhimurium. J Bacteriol. 1985 Feb;161(2):673–680. doi: 10.1128/jb.161.2.673-680.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Săsárman A., Desrochers M. Uroporphyrinogen III cosynthase-deficient mutant of Salmonella typhimurium LT2. J Bacteriol. 1976 Dec;128(3):717–721. doi: 10.1128/jb.128.3.717-721.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Săsărman A., Chartrand P., Lavoie M., Tardif D., Proschek R., Lapointe C. Mapping of a new hem gene in Escherichia coli K12. J Gen Microbiol. 1979 Aug;113(2):297–303. doi: 10.1099/00221287-113-2-297. [DOI] [PubMed] [Google Scholar]
- Săsărman A., Chartrand P., Proschek R., Desrochers M., Tardif D., Lapointe C. Uroporphyrin-accumulating mutant of Escherichia coli K-12. J Bacteriol. 1975 Dec;124(3):1205–1212. doi: 10.1128/jb.124.3.1205-1212.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Săsărman A., Desrochers M., Sonea S., Sanderson K. E., Surdeanu M. Porphobilinogen-accumulating mutants of Salmonella typhimurium LT2. J Gen Microbiol. 1976 Jun;94(2):359–366. doi: 10.1099/00221287-94-2-359. [DOI] [PubMed] [Google Scholar]
- Săsărman A., Sanderson K. E., Surdeanu M., Sonea S. Hemin-deficient mutants of Salmonella typhimurium. J Bacteriol. 1970 May;102(2):531–536. doi: 10.1128/jb.102.2.531-536.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Săsărman A., Surdeanu M., Horodniceanu T. Locus determining the synthesis of delta-aminolevulinic acid in Escherichia coli K-12. J Bacteriol. 1968 Nov;96(5):1882–1884. doi: 10.1128/jb.96.5.1882-1884.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tait G. H. Coproporphyrinogenase activities in extracts of Rhodopseudomonas spheroides and Chromatium strain D. Biochem J. 1972 Aug;128(5):1159–1169. doi: 10.1042/bj1281159. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thomas S. D., Jordan P. M. Nucleotide sequence of the hemC locus encoding porphobilinogen deaminase of Escherichia coli K12. Nucleic Acids Res. 1986 Aug 11;14(15):6215–6226. doi: 10.1093/nar/14.15.6215. [DOI] [PMC free article] [PubMed] [Google Scholar]
- VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
- Verkamp E., Chelm B. K. Isolation, nucleotide sequence, and preliminary characterization of the Escherichia coli K-12 hemA gene. J Bacteriol. 1989 Sep;171(9):4728–4735. doi: 10.1128/jb.171.9.4728-4735.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Way J. C., Davis M. A., Morisato D., Roberts D. E., Kleckner N. New Tn10 derivatives for transposon mutagenesis and for construction of lacZ operon fusions by transposition. Gene. 1984 Dec;32(3):369–379. doi: 10.1016/0378-1119(84)90012-x. [DOI] [PubMed] [Google Scholar]
- Wulff D. L. Delta-aminolevulinic acid-requiring mutant from Escherichia coli. J Bacteriol. 1967 Apr;93(4):1473–1474. doi: 10.1128/jb.93.4.1473-1474.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zagorec M., Buhler J. M., Treich I., Keng T., Guarente L., Labbe-Bois R. Isolation, sequence, and regulation by oxygen of the yeast HEM13 gene coding for coproporphyrinogen oxidase. J Biol Chem. 1988 Jul 15;263(20):9718–9724. [PubMed] [Google Scholar]