title: The characterization and evolutionary relationships of a trypanosomal thiolase.
abstract: Thiolases are enzymes that remove an acetyl-coenzyme A group from acyl-CoA in the catabolic β-oxidation of fatty acids, or catalyse the reverse condensation reaction for anabolic processes such as the biosynthesis of sterols and ketone bodies. In humans, six homologous isoforms of thiolase have been described, differing from each other in sequence, oligomeric state, substrate specificity and subcellular localization. A bioinformatics analysis of parasite genomes, being (i) different species of African trypanosomes, (ii) Trypanosoma cruzi and (iii) Leishmania spp., using the six human sequences as queries, showed that the distribution of thiolases in human and each of the studied Trypanosomatidae is completely different. Only one of these isoforms, called SCP2-thiolase, was found in each of the Trypanosomatidae, whereas the TFE-thiolase was also found in T. cruzi and Leishmania spp., and the AB-thiolase only in T. cruzi. Each of the trypanosomatid thiolases clusters with its orthologues from other organisms in a phylogenetic analysis and shares with them the isoform-specific sequence fingerprints. The single T. brucei SCP2-thiolase has been expressed in Escherichia coli and characterized. It shows activity in both the degradative and synthetic directions. Transcripts of this thiolase were detected in both bloodstream- and procyclic-form trypanosomes, but the protein was found only in the procyclic form. The encoded protein has both a predicted N-terminal mitochondrial signal peptide and a C-terminal candidate type 1 peroxisomal-targeting signal for sorting it into glycosomes. However experimentally, only a mitochondrial localization was found for both procyclic trypanosomes grown with glucose and cells cultured with amino acids as an energy source. When the thiolase expression in procyclic cells was knocked down by RNA interference, no important change in growth rate occurred, irrespective of whether the cells were grown with or without glucose, indicating that the metabolic pathway(s) involving this enzyme is/are not essential for the parasite under either of these growth conditions.
authors: Research Unit for Tropical Diseases, de Duve Institute and Laboratory of Biochemistry, Université catholique de Louvain, TROP 74.39, Avenue Hippocrate 74, B-1200 Brussels, Belgium.
authors: Muriel
authors: M
authors: Mazet
authors: Rajesh K
authors: RK
authors: Harijan
authors: Tiila-Riika
authors: TR
authors: Kiema
authors: Antti M
authors: AM
authors: Haapalainen
authors: Pauline
authors: P
authors: Morand
authors: Jorge
authors: J
authors: Morales
authors: Frédéric
authors: F
authors: Bringaud
authors: Rik K
authors: RK
authors: Wierenga
authors: Paul A M
authors: PA
authors: Michels
mesh_terms: D000102:Acetyl-CoA C-Acyltransferase / Q000235:genetics* / Q000378:metabolism*; D003001:Cloning, Molecular; D016000:Cluster Analysis; D019295:Computational Biology / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D020869:Gene Expression Profiling; D006801:Humans; D007527:Isoenzymes / Q000235:genetics; D007891:Leishmania / Q000201:enzymology; D008928:Mitochondria / Q000201:enzymology; D010802:Phylogeny; D021382:Protein Sorting Signals; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014345:Trypanosoma / Q000201:enzymology*
chemical_list: D007527:Isoenzymes; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D000102:Acetyl-CoA C-Acyltransferase