Probing for primary functions of prohibitin in Trypanosoma brucei.
Prohibitins (PHBs) 1 and 2 are small conserved proteins implicated in a number of functions in the mitochondrion, as well as in the nucleus of eukaryotic cells. The current understanding of PHB functions comes from studies of model organisms such as yeast, worm and mouse, but considerable debate remains with regard to the primary functions of these ubiquitous proteins. We exploit the tractable reverse genetics of Trypanosoma brucei, the causative agent of African sleeping sickness, in order to specifically analyse the function of PHB in this highly divergent eukaryote. Using inducible RNA interference (RNAi) we show that PHB1 is essential in T. brucei, where it is confined to the cell's single mitochondrion forming a high molecular weight complex. PHB1 and PHB2 appear to be indispensible for mitochondrial translation. Their ablation leads to a decrease in mitochondrial membrane potential, however no effect on the level of reactive oxygen species was observed. Flagellates lacking either PHB1 or both PHB1 and PHB2 exhibit significant morphological changes of their organelle, most notably its inflation. Even long after the loss of the PHB proteins, mtDNA was unaltered and mitochondrial cristae remained present, albeit displaced to the periphery of the mitochondrion, which is in contrast to other eukaryotes.
Týc
Jirí
J
Biology Centre, Institute of Parasitology, Czech Academy of Sciences, and Faculty of Natural Sciences, University of South Bohemia, Ceské Budejovice (Budweis), Czech Republic.
Faktorová
Drahomíra
D
Kriegová
Eva
E
Jirků
Milan
M
Vávrová
Zuzana
Z
Maslov
Dmitri A
DA
Lukes
Julius
J
D000818:Animals; D004272:DNA, Mitochondrial / Q000235:genetics / Q000378:metabolism; D008564:Membrane Potentials; D008928:Mitochondria / Q000378:metabolism / Q000648:ultrastructure; D010802:Phylogeny; D000091402:Prohibitins; D014176:Protein Biosynthesis; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D014346:Trypanosoma brucei brucei / Q000235:genetics / Q000254:growth & development / Q000378:metabolism / Q000648:ultrastructure
D004272:DNA, Mitochondrial; C000717333:Phb2 protein, mouse; D000091402:Prohibitins; D015800:Protozoan Proteins; D012333:RNA, Messenger; D012097:Repressor Proteins