title: Downregulation of mitochondrial porin inhibits cell growth and alters respiratory phenotype in Trypanosoma brucei.
abstract: Porin is the most abundant outer membrane (OM) protein of mitochondria. It forms the aqueous channel on the mitochondrial OM and mediates major metabolite flux between mitochondria and cytosol. Mitochondrial porin in Trypanosoma brucei, a unicellular parasitic protozoan and the causative agent of African trypanosomiasis, possesses a beta-barrel structure similar to the bacterial OM porin OmpA. T. brucei porin (TbPorin) is present as a monomer as well as an oligomer on the mitochondrial OM, and its expression is developmentally regulated. In spite of its distinct structure, the TbPorin function is similar to those of other eukaryotic porins. TbPorin RNA interference (RNAi) reduced cell growth in both procyclic and bloodstream forms. The depletion of TbPorin decreased ATP production by inhibiting metabolite flux through the OM. Additionally, the level of trypanosome alternative oxidase (TAO) decreased, whereas the levels of cytochrome-dependent respiratory complexes III and IV increased in TbPorin-depleted mitochondria. Furthermore, the depletion of TbPorin reduced cellular respiration via TAO, which is not coupled with oxidative phosphorylation, but increased the capacity for cyanide-sensitive respiration. Together, these data reveal that TbPorin knockdown reduced the mitochondrial ATP level, which in turn increased the capacity of the cytochrome-dependent respiratory pathway (CP), in an attempt to compensate for the mitochondrial energy crisis. However, a simultaneous decrease in the substrate-level phosphorylation due to TbPorin RNAi caused growth inhibition in the procyclic form. We also found that the expressions of TAO and CP proteins are coordinately regulated in T. brucei according to mitochondrial energy demand.
authors.affiliation: Department of Microbial Pathogenesis and Immune Response, Meharry Medical College, Nashville, TN 37208, USA.
authors.forename: Ujjal K
authors.initials: UK
authors.lastname: Singha
authors.forename: Shvetank
authors.initials: S
authors.lastname: Sharma
authors.forename: Minu
authors.initials: M
authors.lastname: Chaudhuri
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019069:Cell Respiration; D015536:Down-Regulation; D018507:Gene Expression Regulation, Developmental; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D014346:Trypanosoma brucei brucei / Q000737:chemistry / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D050994:Voltage-Dependent Anion Channels / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D015800:Protozoan Proteins; D050994:Voltage-Dependent Anion Channels; D010088:Oxidoreductases; C020094:duroquinol oxidase