Autophagy in Trypanosoma brucei: amino acid requirement and regulation during different growth phases.
Autophagy in the protozoan parasite, Trypanosoma brucei, may be involved in differentiation between different life cycle forms and during growth in culture. We have generated multiple parasite cell lines stably expressing green fluorescent protein- or hemagglutinin-tagged forms of the autophagy marker proteins, TbAtg8.1 and TbAtg8.2, in T. brucei procyclic forms to establish a trypanosome system for quick and reliable determination of autophagy under different culture conditions using flow cytometry. We found that starvation-induced autophagy in T. brucei can be inhibited by addition of a single amino acid, histidine, to the incubation buffer. In addition, we show that autophagy is induced when parasites enter stationary growth phase in culture and that their capacity to undergo starvation-induced autophagy decreases with increasing cell density.
Institute of Biochemistry and Molecular Medicine, University of Bern, Bern, Switzerland; Graduate School for Cellular and Biomedical Sciences, University of Bern, Bern, Switzerland.
Remo S
RS
Schmidt
Institute of Biochemistry and Molecular Medicine, University of Bern, Bern, Switzerland.
Peter
P
Bütikofer
D000596:Amino Acids / Q000378:metabolism*; D001343:Autophagy / Q000502:physiology*; D002454:Cell Differentiation; D016054:DNA, Protozoan / Q000378:metabolism; D015800:Protozoan Proteins / Q000378:metabolism*; D014346:Trypanosoma brucei brucei / Q000254:growth & development / Q000378:metabolism*
D000596:Amino Acids; D016054:DNA, Protozoan; D015800:Protozoan Proteins