Cytochrome c550 expression in Paracoccus denitrificans strongly depends on growth condition: identification of promoter region for cycA by transcription start analysis
The periplasmic cytochrome c550 content of Paracoccus denitrificans has been shown by immunological detection to be strongly dependent on the mode of growth. Cells grown under anaerobic, denitrifying conditions or methylotrophically in the presence of oxygen contained substantially more cytochrome c550 than cells grown aerobically on multicarbon substrates. A similar pattern was observed when expression of the cycA gene (encoding cytochrome c550), was monitored using an Escherichia coli alkaline phosphatase gene (phoA) fusion as a reporter of cycA promoter activity. The increase in cycA expression observed during growth on C1 substrates was substantially diminished if succinate was also present. These results reveal that expression of cycA is subject to multiple regulatory controls and suggest that cytochrome c550 has a general role in electron transfer to periplasmic reductases required for anaerobic denitrifying growth and from dehydrogenases required for aerobic growth on C1 compounds. Two major transcriptional initiation start points for the cycA gene have been identified.
AlefounderP. R., FergusonS. J.1981; A periplasmic location for methanol dehydrogenase from Paraeoccus denitrificans: implications for proton pumping by cytochrome aa3. Biochem Biophys Res Commun 98:778–784
BagdasarianM., LurzR., RückertB., FranklinF. C. H., BagdasarianM. M., FreyJ., TimmisK. N.1981; Specific-purpose plasmid cloning vector II. Broad host range, high copy number RSFl0l0-derived vectors and a host-vector system for cloning in Pseudomonas. Gene 16:237–247
BenningM. M., MeyerT. E., HoldenH. M.1994; X-ray structure of the cytochrome c2 isolated from Paraeoccus denitrificansrefined to 1·7 Å resolution. Arch Biochem Biophys 310:460–466
BerksB. C., BarattaD., RichardsonJ., FergusonS. J.1993; Purification and characterisation of a nitrous oxide reductase from Thiosphaera pantotropha. Implications for the mechanism of aerobic nitrous oxide reduction. Eur J Biochem 212:467–476
de BoerA. P. N., ReijndersW. N. M., KuenenJ. G., StouthamerA. H., van SpanningR. J. M.1994; Isolation, sequencing and mutational analysis of a gene cluster involved in nitrite reduction in Paraeoccus denitrificans. Antonie Leeuwenhoek 66:111–127
BosmaG., BrasterM., StouthamerA. H., van VerseveldH. W.1987; Subfractionation and characterisation of soluble c-type cytochromes from Paraeoccus denitrificans cultured under various limiting conditions in the chemosrat. Eur J Biochem 165:665–670
BottM., Thöny-MeyerL, LofererH., RossbachS., TullyR. E., KeisterD., ApplebyC. A., HenneckeH.1995; Bradyrhizobium japonicum cytochrome c550 is required for nitrate respiration but not for symbiotic nitrogen fixation. J Bacteriol 177:2214–2217
BradfordM. M.1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
BrickmanE., BeckwithJ.1975; Analysis of the regulation of Escherichia coli alkaline phosphatase synthesis using deletions and ɸ80 transducing phages. J Mol Biol 96:307–316
BurnellJ. N., JohnP., WhatleyF. R.1975; The reversibility of active sulphate transport in membrane vesicles of Paraeoccus denitrificans. Biochem J 150:527–536
DavidsonV. L., KumarM. A.1989; Cytochrome c550 mediates electron transfer from inducible periplasmic cytochromes to the cytoplasmic membrane of Paraeoccus denitrificans. FEBS Lett 245:271–273
DickinsM. A., DhawanT., GunsalusR. P., SchroederI., CecchiniG.1995; Cloning, sequencing and expression of the succinate-ubiquinone oxidoreductase (SdhCDAB) operon from Paraeoccus denitrificans. EMBL database entry PDU31902.
ErasoJ. M., KaplanS.1994; prr4, a putative response regulator involved in oxygen regulation of photosynthesis gene expression in Rhodobacter sphaeroides. J Bacterial 176:32–43
FergusonS. J., PageM. D.1990; The functions and biosynthesis of the c-type cytochromes of bacterial respiratory chains with particular reference to autotrophs and the handling of C1compounds. FEMS Microbiol Rev 87:227–234
FigurskiD. H., HelinskiD. R.1979; Replication of an origin- containing derivative of plasmid RK2 dependent on a plasmid function derived in trans. Proc Natl Acad Sci USA761648–1652
de GierJ. W., LubbenM., ReijndersW. N., TipkerC. A., SlotboomD. J., van SpanningR. J., StouthamerA. H., van der OostJ.1994; The terminal oxidases of Paraeoccus denitrificans. Mol Microbiol 13:183–196
De GierJ. L., van der OostJ., HarmsN., StouthamerA. H., van SpanningR.J.M. 1995; The oxidation of methylamine in Paraeoccus denitrificans. Eur J Biochem 229:148–154
GoodhewC. F., BrownK. R., PettigrewG. W.1986; Haem staining in gels, a useful tool in the study of Bacteriol c-type cytochromes. Biochim Biophys Acta 852:288–294
GoodhewC. F., PettigrewG. W., DevreeseB., van BeeumenJ., van SpanningR. J. M., BakerS. C., SaundersN., FergusonS. J., ThompsonI. P.1996; The cytochromes c550 of Paraeoccus denitrificans and Thiosphaera pantotropha: a need for re-evaluation of the history of Paraeoccus cultures. FEMS Microbiol Lett 137:95–101
HarmsN., ReijndersW. N. M., AnazawaH., van der PalenC.J.N.M., van SpanningR. J. M., OltmanL. F., StouthamerA. H.1993; Identification of a two-component regulatory system controlling methanol dehydrogenase synthesis in Paraeoccus denitrificans. Mol Microbiol 8:457–470
KimK.S, GuarenteL.1989; Mutations that alter transcriptional activation but not DNA binding in the zinc finger of yeast activator HAP1. Nature 342:200–203
MatchovaI., KuceraI.1991; Evidence for the role of soluble cytochrome c in the dissimilatory reduction of nitrite and nitrous oxide by cells of Paracoccus denitrificans. Biochim Biophys Acta 1058:256–260
MoirJ. W. B., FergusonS. J.1994; Properties of a Paracoccus denitrificans mutant deleted in cytochrome c550 indicate that a copper protein can substitute for this cytochrome in electron transport to nitrite, nitric oxide and nitrous oxide. Microbiology 140:389–397
MoirJ. W. B., BarattaD., RichardsonD. J., FergusonS. J.1993; The purification of a cd1-type nitrite reductase and the absence of a copper type nitrite reductase from the aerobic denitrifier Thiosphaera pantotropha; the role of pseudoazurin as an electron donor. Eur J Biochem 212:377–385
MoirJ. W. B., RichardsonD. J., FergusonS. J.1995; The expression of redox proteins of denitrification in Thiosphaera pantotropha grown with oxygen, nitrate and nitrous oxide. Arch Microbiol 164:43–49
PageM. D., FergusonS. J.1990; Apo-forms of cytochrome c550 and cytochrome cd1 are translocated to the periplasm of Paracoccus denitrificans in the absence of haem incorportion caused by either mutation or inhibition of haem synthesis. Mol Microbiol 4:1181–1192
PageM. D., FergusonS. J.1993; Mutants of Methylobacterium extorquens and Paracoccus denitrificans deficient in c-type cytochrome biogenesis synthesise the methylamine-dehydrogenase polypeptides that cannot assemble the tryptophan-tryptophylquinone group. Eur J Biochem 218:711–717
PageM. D., FergusonS. J.1994; Differential reduction in soluble and membrane-bound cytochrome contents in a Paracoccus denitrificans mutant partially deficient in 5-aminolevulinate synthase activity. J Bacteriol 176:5919–5928
PageM. D., FergusonS. J.1995; Cloning and sequence analysis of cycH gene from Paracoccus denitrificans; the cycH gene product is required for assembly of all c-type cytochromes, including cytochrome c1. Mol Microbiol 15:307–315
PageM. D., CarrG., BellL. C., FergusonS. J.1989; Structure, control and assembly of a Bacteriol electron transport system as exemplified by Paracoccus denitrificans. Biochem Soc Trans 17:991–993
ParsonageD., GreenfieldA. J., FergusonS. J.1986; Evidence that the energy conserving electron transport pathways to nitrate and cytochrome o branch at ubiquinone in Paracoccus denitrificans. Arch Microbiol 145:191–196
RichardsonD. J., BellL. C., McewanA. G., JacksonJ. B., FergusonS. J.1991; Cytochrome c2 is essential for electron transfer to nitrous oxide reductase from physiological substrates in Rhodobacter capsulatus and can act as an electron donor to the reductase in vitro. Eur J Biochem 199:677–683
RichterO.-M.H., TaoJ.-S., TurbaA., LudwigB.1994; A cytochrome ba3 functions as a quinol oxidase in Paracoccus denitrificans. Purification, cloning and sequence comparison. J Biol Chem 269:23079–23086
SambongiY., FergusonS. J.1994; Synthesis of holo Paracoccus denitrificans cytochrome c550 requires targeting to the periplasm whereas that of holo Hydrogenobacter thermcphilus cytochrome c552 does not. FEBS Lett 340:65–70
SambongiY., StollR., FergusonS. J.1996; Alteration of haem attachment and signal cleavage sites for Paracoccus denitrificanscytochrome c550 probes pathway of c-type cytochrome biogenesis in Escherichia coli. Mol Microbiol 19:1193–1204
SamynB., BerksB. C., PageM. D., FergusonS. J., van BeeumenJ. J.1994; Characterisation and amino acid sequence of cytochrome c550 from Thiosphaera pantotropha. Eur J Biochem 219:585–594
SapsheadL. M., WimpennyJ. W. T.1972; The influence of oxygen and nitrate on the formation of the cytochrome pigments of the aerobic and anaerobic respiratory chain of Micrococcus denitrificans. Biochim Biophys Acta 267:388–397
ScholesP. B., SmithL.1968; Composition and properties of the membrane-bound respiratory chain system of Micrococcus denitrificans. Biochim Biophys Acta 153:363–375
van SpanningR. J. M., WansellW. N. M., HarmsN., RasJ., OltmannL. F., StouthamerA. H.1990; Mutagenesis of the gene encoding cytochrome c550 of Paracoccus denitrificans and analysis of the resultant physiological effects. J Bacteriol 172:986–996
van SpanningR. J. M., WansellC. W., ReijndersW. N. M., HarmsN., RasJ., OltmannL. F., StouthamerA. H.1991; A method for introduction of unmarked mutations in the genome of Paracoccus denitrificans: construction of strains with multiple mutations in the genes encoding periplasmic cytochromes c550, c5511and c553i. J Bacteriol 173:6962–6970
van SpanningR. J. M., van der PalenC. J. N. M., SlotboomD.-J., ReijndersW. N. M., StouthamerA. H., DuineJ. A.1995a; Expression of the mau genes involved in methylamine metabolism in Paracoccus denitrificans is under control of a LysR-type transcriptional activator. Eur J Biochem 226:201–210
van SpanningR. J. M., de BoerA. P. N., ReijndersW. N. M., SpiroS., WesterhoffH. V., StouthamerA. H., van der OostJ.1995b; Nitrite and nitric oxide reduction in Paracoccus denitrificansis under the control of NNR, a regulatory protein that belongs to the FNR family of transcriptional activators. FEBS Lett 360:151–154
TzagoloffA., CapitanioN., NobregaM. P., GattiD.1990; Cytochrome oxidase assembly in yeast requires the product of COX11, a homolog of the P. denitrificans protein encoded by ORF3. EMBO J 9:2759–2764
de VriesG. E., HarmsN., HoogendijkJ., StoutharnerA. H.1989; Isolation and characterisation of Paracoccus denitrificans mutants with increased conjugation frequencies and pleiotropic loss of a (nGATCn) DNA modifying property. Arch Microbiol 152:52–57
Cytochrome c550 expression in Paracoccus denitrificans strongly depends on growth condition: identification of promoter region for cycA by transcription start analysis