Abstract
An Escherichia coli mutant with a disrupted visA gene was defective in ferrochelatase activity but expressed wild-type levels of protoporphyrinogen oxidase activity. The visA coding region was placed under the transcriptional control of T7 RNA polymerase in an E. coli expression system, and the product was expressed as a 38-kDa protein. The overexpressed protein was purified to near homogeneity and was found to contain ferrochelatase activity. The data show that the visA gene encodes ferrochelatase, and we propose that it be renamed hemH to reflect that conclusion.
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- Alefounder P. R., Abell C., Battersby A. R. The sequence of hemC, hemD and two additional E. coli genes. Nucleic Acids Res. 1988 Oct 25;16(20):9871–9871. doi: 10.1093/nar/16.20.9871. [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]
- Brenner D. A., Didier J. M., Frasier F., Christensen S. R., Evans G. A., Dailey H. A. A molecular defect in human protoporphyria. Am J Hum Genet. 1992 Jun;50(6):1203–1210. [PMC free article] [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]
- 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]
- Ferreira G. C., Andrew T. L., Karr S. W., Dailey H. A. Organization of the terminal two enzymes of the heme biosynthetic pathway. Orientation of protoporphyrinogen oxidase and evidence for a membrane complex. J Biol Chem. 1988 Mar 15;263(8):3835–3839. [PubMed] [Google Scholar]
- Frustaci J. M., O'Brian M. R. Characterization of a Bradyrhizobium japonicum ferrochelatase mutant and isolation of the hemH gene. J Bacteriol. 1992 Jul;174(13):4223–4229. doi: 10.1128/jb.174.13.4223-4229.1992. [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]
- Ilag L. L., Jahn D., Eggertsson G., Söll D. The Escherichia coli hemL gene encodes glutamate 1-semialdehyde aminotransferase. J Bacteriol. 1991 Jun;173(11):3408–3413. doi: 10.1128/jb.173.11.3408-3413.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jahn D., Michelsen U., Söll D. Two glutamyl-tRNA reductase activities in Escherichia coli. J Biol Chem. 1991 Feb 5;266(4):2542–2548. [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]
- Labbe-Bois R. The ferrochelatase from Saccharomyces cerevisiae. Sequence, disruption, and expression of its structural gene HEM15. J Biol Chem. 1990 May 5;265(13):7278–7283. [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]
- Li J. M., Umanoff H., Proenca R., Russell C. S., Cosloy S. D. Cloning of the Escherichia coli K-12 hemB gene. J Bacteriol. 1988 Feb;170(2):1021–1025. doi: 10.1128/jb.170.2.1021-1025.1988. [DOI] [PMC free article] [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]
- Miyamoto K., Nishimura K., Masuda T., Tsuji H., Inokuchi H. Accumulation of protoporphyrin IX in light-sensitive mutants of Escherichia coli. FEBS Lett. 1992 Oct 5;310(3):246–248. doi: 10.1016/0014-5793(92)81341-i. [DOI] [PubMed] [Google Scholar]
- Nakahashi Y., Fujita H., Taketani S., Ishida N., Kappas A., Sassa S. The molecular defect of ferrochelatase in a patient with erythropoietic protoporphyria. Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):281–285. doi: 10.1073/pnas.89.1.281. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nakahigashi K., Nishimura K., Miyamoto K., Inokuchi H. Photosensitivity of a protoporphyrin-accumulating, light-sensitive mutant (visA) of Escherichia coli K-12. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10520–10524. doi: 10.1073/pnas.88.23.10520. [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]
- Rosenberg A. H., Lade B. N., Chui D. S., Lin S. W., Dunn J. J., Studier F. W. Vectors for selective expression of cloned DNAs by T7 RNA polymerase. Gene. 1987;56(1):125–135. doi: 10.1016/0378-1119(87)90165-x. [DOI] [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]
- Siepker L. J., Kramer S. Protoporphyrin accumulation by mitogen stimulated lymphocytes and protoporphyrinogen oxidase activity in patients with porphyria variegata and erythropoietic protoporphyria: evidence for deficiency of protoporphyrinogen oxidase and ferrochelatase in both diseases. Br J Haematol. 1985 May;60(1):65–74. doi: 10.1111/j.1365-2141.1985.tb07386.x. [DOI] [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]
- Taketani S., Nakahashi Y., Osumi T., Tokunaga R. Molecular cloning, sequencing, and expression of mouse ferrochelatase. J Biol Chem. 1990 Nov 15;265(32):19377–19380. [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]
- 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]


