Cytochrome oxidase subunit VI of Trypanosoma brucei is imported without a cleaved presequence and is developmentally regulated at both RNA and protein levels.
Mitochondrial respiration in the African trypanosome undergoes dramatic developmental stage regulation. This requires co-ordinated control of components encoded by both the nuclear genome and the kinetoplast, the unusual mitochondrial genome of these parasites. As a model for understanding the co-ordination of these genomes, we have examined the regulation and mitochondrial import of a nuclear-encoded component of the cytochrome oxidase complex, cytochrome oxidase subunit VI (COXVI). By generating transgenic trypanosomes expressing intact or mutant forms of this protein, we demonstrate that COXVI is not imported using a conventional cleaved presequence and show that sequences at the N-terminus of the protein are necessary for correct mitochondrial sorting. Analyses of endogenous and transgenic COXVI mRNA and protein expression in parasites undergoing developmental stage differentiation demonstrates a temporal order of control involving regulation in the abundance of, first, mRNA and then protein. This represents the first dissection of the regulation and import of a nuclear-encoded protein into the cytochrome oxidase complex in these organisms, which were among the earliest eukaryotes to possess a mitochondrion.
Tasker
M
M
School of Biological Sciences, 2.14 Stopford Building, University of Manchester, Oxford Road, Manchester UK M13 9PT, UK.
Timms
M
M
Hendriks
E
E
Matthews
K
K
D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D002458:Cell Fractionation; D003470:Culture Media; D003576:Electron Transport Complex IV / Q000235:genetics* / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D018507:Gene Expression Regulation, Developmental; D008928:Mitochondria / Q000201:enzymology; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D016053:RNA, Protozoan / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D016833:Trypanosoma brucei rhodesiense / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development
D003470:Culture Media; D015800:Protozoan Proteins; D016053:RNA, Protozoan; D003576:Electron Transport Complex IV