title: Identification and validation of Trypanosoma cruzi's glycosomal adenylate kinase containing a peroxisomal targeting signal.
abstract: Adenylate kinases are key enzymes involved in cell energy management. Trypanosomatid organisms have the largest number of isoforms found in a single cell, constituting a major difference with the mammalian hosts. In this work we study an adenylate kinase, TcADK3, the only Trypanosoma cruzi protein harboring the putative peroxisomal (glycosomal) targeting signal, "-CKL". Parasites expressing GFP fused to TcADK3 showed a strong fluorescence in the glycosomes. The same result was obtained when the tripeptide "-CKL" was added at the C-terminus of the GFP, demonstrating that this signal is necessary and sufficient for targeting proteins to glycosomes. When this tripeptide was removed from the GFP-TcADK3 fusion protein, the fluorescence was re-localized in the cytoplasm. The CKL signal could be used for targeting foreign proteins to the glycosomes. This model also provides a useful tool to study glycosomes dynamics, morphology or number in living parasites in any stage of the life cycle.
authors.affiliation: Laboratorio de Biología Molecular de Trypanosoma cruzi (LBMTC), Instituto de Investigaciones Médicas Alfredo Lanari, Universidad de Buenos Aires and CONICET, Buenos Aires, Argentina.
authors.forename: María de Los
authors.initials: Mde L
authors.lastname: Milagros Camara
authors.forename: León A
authors.initials: LA
authors.lastname: Bouvier
authors.forename: Mariana R
authors.initials: MR
authors.lastname: Miranda
authors.forename: Claudio A
authors.initials: CA
authors.lastname: Pereira
mesh_terms: D000263:Adenylate Kinase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D015971:Gene Expression Regulation, Enzymologic; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008830:Microbodies / Q000201:enzymology*; D008856:Microscopy, Fluorescence; D020675:Peroxisomes / Q000201:enzymology*; D010957:Plasmids; D015398:Signal Transduction; D014162:Transfection; D014349:Trypanosoma cruzi / Q000201:enzymology* / Q000235:genetics
chemical_list: D007527:Isoenzymes; D000263:Adenylate Kinase