Import of a constitutively expressed protein into mitochondria from procyclic and bloodstream forms of Trypanosoma brucei.

Trypanosoma brucei developmentally regulates mitochondrial function during its life cycle. Numerous nuclear encoded mitochondrial proteins undergo posttranslational regulation in a developmental fashion, but exactly how that regulation is achieved is unclear. We are interested in mitochondrial import as a potential regulatory step for nuclear encoded mitochondrial proteins. Previously, an in vitro import system was developed for the procyclic lifestage. We report here the development of an in vitro import system for bloodstream trypanosomes using a crude mitochondrial preparation. NADH dehydrogenase subunit K (NdhK) is a nuclear encoded mitochondrial protein that is constitutively expressed in bloodstream and procyclic trypanosomes. We examined the import of NdhK into procylic and bloodstream mitochondria in vitro. In both lifestages import of NdhK requires a membrane potential across the inner mitochondrial membrane, mitochondrial matrix ATP, and is time dependent. The precursor protein is processed by a matrix associated metalloprotease in a single cleavage step to mature protein.