The Escherichia Coli CcmG Protein Fulfils a Specific Role in Cytochrome C Assembly

Overview

Journal Biochem J

Specialty Biochemistry

Date 2001 Mar 21

PMID 11256948

Citations 26

Authors

E Reid

J Cole

D J Eaves

Affiliations

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Abstract

In Escherichia coli K-12, c-type cytochromes are synthesized only during anaerobic growth with trimethylamine-N-oxide, nitrite or low concentrations of nitrate as the terminal electron acceptor. A thioredoxin-like protein, CcmG, is one of 12 proteins required for their assembly in the periplasm. Its postulated function is to reduce disulphide bonds formed between correctly paired cysteine residues in the cytochrome c apoproteins prior to haem attachment by CcmF and CcmH. We report that loss of CcmG synthesis by mutation was not compensated by a second mutation in disulphide-bond-forming proteins, DsbA or DsbB, or by the chemical reductant, 2-mercaptoethanesulphonic acid. An anti-CcmG polyclonal antibody was used in Western-blot analysis to probe the redox state of CcmG in mutants defective in the synthesis of other proteins essential for cytochrome c assembly. The oxidized form of CcmG accumulated not only in trxA or dipZ mutants defective in the transfer of electrons from the cytoplasm for disulphide isomerization and reduction reactions in the periplasm, but also in ccmF and ccmH mutants. The requirement of both CcmF and CcmH for the reduction of the disulphide bond in CcmG indicates that CcmG functions later than CcmF and CcmH in cytochrome c assembly, rather than in electron transfer from the membrane-associated DipZ (also known as DsbD) to CcmH. The data support a model proposed by others in which CcmG catalyses one of the last reactions specific to cytochrome c assembly.

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References

Mejean V, Lepelletier M, Giordano G, Chippaux M, PASCAL M . TMAO anaerobic respiration in Escherichia coli: involvement of the tor operon. Mol Microbiol. 1994; 11(6):1169-79. DOI: 10.1111/j.1365-2958.1994.tb00393.x. View

Thony-Meyer L, Hederstedt L . Escherichia coli ccm in-frame deletion mutants can produce periplasmic cytochrome b but not cytochrome c. FEBS Lett. 1997; 410(2-3):351-5. DOI: 10.1016/s0014-5793(97)00656-x. View

Hussain H, Grove J, Griffiths L, Busby S, Cole J . A seven-gene operon essential for formate-dependent nitrite reduction to ammonia by enteric bacteria. Mol Microbiol. 1994; 12(1):153-63. DOI: 10.1111/j.1365-2958.1994.tb01004.x. View

Sambongi Y, Ferguson S . 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; 353(3):235-8. DOI: 10.1016/0014-5793(94)01053-6. View

Crooke H, Cole J . 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; 15(6):1139-50. DOI: 10.1111/j.1365-2958.1995.tb02287.x. View

Stewart E, KATZEN F, Beckwith J . Six conserved cysteines of the membrane protein DsbD are required for the transfer of electrons from the cytoplasm to the periplasm of Escherichia coli. EMBO J. 1999; 18(21):5963-71. PMC: 1171662. DOI: 10.1093/emboj/18.21.5963. View

Chung J, Chen T, Missiakas D . Transfer of electrons across the cytoplasmic membrane by DsbD, a membrane protein involved in thiol-disulphide exchange and protein folding in the bacterial periplasm. Mol Microbiol. 2000; 35(5):1099-109. DOI: 10.1046/j.1365-2958.2000.01778.x. View

Gordon E, Page M, Willis A, Ferguson S . Escherichia coli DipZ: anatomy of a transmembrane protein disulphide reductase in which three pairs of cysteine residues, one in each of three domains, contribute differentially to function. Mol Microbiol. 2000; 35(6):1360-74. DOI: 10.1046/j.1365-2958.2000.01796.x. View

Thony-Meyer L . Biogenesis of respiratory cytochromes in bacteria. Microbiol Mol Biol Rev. 1997; 61(3):337-76. PMC: 232615. DOI: 10.1128/mmbr.61.3.337-376.1997. View

10.

Monika E, Goldman B, Beckman D, Kranz R . A thioreduction pathway tethered to the membrane for periplasmic cytochromes c biogenesis; in vitro and in vivo studies. J Mol Biol. 1997; 271(5):679-92. DOI: 10.1006/jmbi.1997.1227. View

11.

Raina S, Missiakas D . Making and breaking disulfide bonds. Annu Rev Microbiol. 1997; 51:179-202. DOI: 10.1146/annurev.micro.51.1.179. View

12.

Kobayashi T, Kishigami S, Sone M, Inokuchi H, Mogi T, Ito K . Respiratory chain is required to maintain oxidized states of the DsbA-DsbB disulfide bond formation system in aerobically growing Escherichia coli cells. Proc Natl Acad Sci U S A. 1997; 94(22):11857-62. PMC: 23636. DOI: 10.1073/pnas.94.22.11857. View

13.

Rietsch A, Bessette P, Georgiou G, Beckwith J . Reduction of the periplasmic disulfide bond isomerase, DsbC, occurs by passage of electrons from cytoplasmic thioredoxin. J Bacteriol. 1997; 179(21):6602-8. PMC: 179585. DOI: 10.1128/jb.179.21.6602-6608.1997. View

14.

Fabianek R, Hennecke H, Thony-Meyer L . The active-site cysteines of the periplasmic thioredoxin-like protein CcmG of Escherichia coli are important but not essential for cytochrome c maturation in vivo. J Bacteriol. 1998; 180(7):1947-50. PMC: 107112. DOI: 10.1128/JB.180.7.1947-1950.1998. View

15.

Page M, Sambongi Y, Ferguson S . Contrasting routes of c-type cytochrome assembly in mitochondria, chloroplasts and bacteria. Trends Biochem Sci. 1998; 23(3):103-8. DOI: 10.1016/s0968-0004(98)01173-6. View

16.

Eaves D, Grove J, Staudenmann W, James P, Poole R, White S . Involvement of products of the nrfEFG genes in the covalent attachment of haem c to a novel cysteine-lysine motif in the cytochrome c552 nitrite reductase from Escherichia coli. Mol Microbiol. 1998; 28(1):205-16. DOI: 10.1046/j.1365-2958.1998.00792.x. View

17.

Zeng H, Snavely I, Zamorano P, Javor G . Low ubiquinone content in Escherichia coli causes thiol hypersensitivity. J Bacteriol. 1998; 180(14):3681-5. PMC: 107339. DOI: 10.1128/JB.180.14.3681-3685.1998. View

18.

Schulz H, Hennecke H, Thony-Meyer L . Prototype of a heme chaperone essential for cytochrome c maturation. Science. 1998; 281(5380):1197-200. DOI: 10.1126/science.281.5380.1197. View

19.

Kranz R, Lill R, Goldman B, Bonnard G, Merchant S . Molecular mechanisms of cytochrome c biogenesis: three distinct systems. Mol Microbiol. 1998; 29(2):383-96. DOI: 10.1046/j.1365-2958.1998.00869.x. View

20.

Reid E, Eaves D, Cole J . The CcmE protein from Escherichia coli is a haem-binding protein. FEMS Microbiol Lett. 1998; 166(2):369-75. DOI: 10.1111/j.1574-6968.1998.tb13914.x. View