An expanded adenylate kinase gene family in the protozoan parasite Trypanosoma cruzi.
Adenylate kinases supply energy routes in cellular energetic homeostasis. In this work, we identified and characterized the adenylate kinase activity in extracts from the flagellated parasite Trypanosoma cruzi, the causative agent of Chagas' disease. Adenylate kinase activity was detected in different subcellular fractions and the cytosolic isoform was biochemically characterized. Cytosolic adenylate kinase specific activity increases continuously during the epimastigote growth and is down-regulated when other soluble phosphotransferase, arginine kinase, is overexpressed. Six different genes of adenylate kinase isoforms were identified and the mRNA expression was confirmed by RT-PCR and Northern Blot. Three open reading frames coding for different enzyme isoforms named TzADK1, TzADK2 and TzADK5 were cloned and functionally expressed in E. coli. This work reports an unusually large number of genes of adenylate kinases and suggests a coordinated regulation of phosphotransferase-mediated ATP regenerating pathways in the unicellular parasite Trypanosoma cruzi.
Departamento de Sustancias Vasoactivas, Instituto de Investigaciones Médicas Alfredo Lanari, Consejo Nacional de Investigaciones Científicas y Técnicas, Universidad de Buenos Aires, Combatientes de Malvinas, Buenos Aires, Argentina.
Leon A
LA
Bouvier
Mariana R
MR
Miranda
Gaspar E
GE
Canepa
Maria Júlia M
MJ
Alves
Claudio A
CA
Pereira
D000263:Adenylate Kinase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D003600:Cytosol / Q000201:enzymology; D004072:Digitonin; D015971:Gene Expression Regulation, Enzymologic; D017125:Genes, Protozoan / Q000235:genetics; D018503:Genome, Protozoan / Q000235:genetics; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D005810:Multigene Family / Q000235:genetics*; D010271:Parasites / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D021381:Protein Transport; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D012995:Solubility; D013347:Subcellular Fractions; D014349:Trypanosoma cruzi / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development
D007527:Isoenzymes; D012333:RNA, Messenger; D000263:Adenylate Kinase; D004072:Digitonin