title: A novel component of the disulfide-reducing pathway required for cytochrome c assembly in plastids.
abstract: In plastids, the conversion of energy in the form of light to ATP requires key electron shuttles, the c-type cytochromes, which are defined by the covalent attachment of heme to a CXXCH motif. Plastid c-type cytochrome biogenesis occurs in the thylakoid lumen and requires a system for transmembrane transfer of reductants. Previously, CCDA and CCS5/HCF164, found in all plastid-containing organisms, have been proposed as two components of the disulfide-reducing pathway. In this work, we identify a small novel protein, CCS4, as a third component in this pathway. CCS4 was genetically identified in the green alga Chlamydomonas reinhardtii on the basis of the rescue of the ccs4 mutant, which is blocked in the synthesis of holoforms of plastid c-type cytochromes, namely cytochromes f and c(6). Although CCS4 does not display sequence motifs suggestive of redox or heme-binding function, biochemical and genetic complementation experiments suggest a role in the disulfide-reducing pathway required for heme attachment to apoforms of cytochromes c. Exogenous thiols partially rescue the growth phenotype of the ccs4 mutant concomitant with recovery of holocytochrome f accumulation, as does expression of an ectopic copy of the CCDA gene, encoding a trans-thylakoid transporter of reducing equivalents. We suggest that CCS4 might function to stabilize CCDA or regulate its activity.
authors.affiliation: Department of Molecular Genetics, The Ohio State University, Columbus, Ohio 43210, USA.
authors.forename: Stéphane T
authors.initials: ST
authors.lastname: Gabilly
authors.forename: Janette
authors.initials: J
authors.lastname: Kropat
authors.forename: Mohamed
authors.initials: M
authors.lastname: Karamoko
authors.forename: M Dudley
authors.initials: MD
authors.lastname: Page
authors.forename: Stacie S
authors.initials: SS
authors.lastname: Nakamoto
authors.forename: Sabeeha S
authors.initials: SS
authors.lastname: Merchant
authors.forename: Patrice P
authors.initials: PP
authors.lastname: Hamel
mesh_terms: D000595:Amino Acid Sequence; D016825:Chlamydomonas reinhardtii / Q000235:genetics* / Q000378:metabolism*; D002736:Chloroplasts / Q000235:genetics / Q000378:metabolism; D003574:Cytochrome c Group / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*; D045304:Cytochromes c / Q000235:genetics* / Q000378:metabolism*; D045348:Cytochromes f / Q000235:genetics / Q000378:metabolism; D004220:Disulfides / Q000378:metabolism; D006418:Heme / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism; D010084:Oxidation-Reduction; D010788:Photosynthesis / Q000235:genetics*; D018087:Plastids / Q000235:genetics / Q000378:metabolism; D020524:Thylakoids / Q000235:genetics / Q000378:metabolism
chemical_list: D003574:Cytochrome c Group; D004220:Disulfides; D050505:Mutant Proteins; D006418:Heme; D045304:Cytochromes c; D045348:Cytochromes f