Navegar

Colección

Datos Estadísticas

Artículo

Estamos trabajando para incorporar este artículo al repositorio
Consulte el artículo en la página del editor
Consulte la política de Acceso Abierto del editor

Abstract:

Trypanosoma cruzi adenylyl cyclases are encoded by a large polymorphic gene family. Although several genes have been identified in this parasite, little is known about the properties and regulation of these enzymes. Here we report the cloning and characterization of TczAC, a novel member of T. cruzi adenylyl cyclase family. The TczAC gene is expressed in all of the parasite life forms and encodes a 1,313-amino acid protein that can complement a Saccharomyces cerevisiae mutant deficient in adenylyl cyclase activity. The recombinant enzyme expressed in yeasts is constitutively active, has a low affinity for ATP (Km = 406 μM), and requires a divalent cation for catalysis. TczAC is inhibited by Zn2+ and the P-site inhibitor 2′-deoxyadenosine 3′-monophosphate, suggesting some level of conservation in the catalytic mechanism with mammalian adenylyl cyclases. It shows a dose-dependent stimulation by Ca2+ which can be reversed by high concentrations of phenothiazinic calmodulin inhibitors. However, bovine calmodulin fails to stimulate the enzyme. Using a yeast two-hybrid screen it was found that TczAC interacts through its catalytic domain with the paraflagellar rod protein, a component of the flagellar structure. Furthermore, we demonstrate that TczAC can dimerize through the same domain. These results provide novel evidence of the possible localization and regulation of this protein.

Registro:

Documento: Artículo
Título:A novel calcium-stimulated adenylyl cyclase from Trypanosoma cruzi, which interacts with the structural flagellar protein paraflagellar rod
Autor:D'Angelo, M.A.; Montagna, A.E.; Sanguineti, S.; Torres, H.N.; Flawiá, M.M.
Filiación:Instituto de Investigaciones en Ingeniería Genética y Biología Molecular, Consejo Nacional de Investigaciones Científicas y Técnicas, Universidad de Buenos Aires, 1428, Argentina
INGEBI, Vuelta de Obligado 2490, Buenos Aires 1428, Argentina
Palabras clave:Amino acids; Calcium; Catalysis; Characterization; Cloning; Dimerization; Enzymes; Genes; Yeast; Dose-dependent stimulation; Biochemistry; 2' deoxyadenosine 3' phosphate; adenosine triphosphate; adenylate cyclase; calmodulin inhibitor; gene product; paraflagellar rod protein; protein TczAC; protozoal protein; unclassified drug; zinc; article; binding affinity; controlled study; enzyme active site; enzyme activity; enzyme inhibition; enzyme regulation; gene expression; genetic code; inhibition kinetics; molecular cloning; multigene family; nonhuman; nucleotide sequence; priority journal; protein localization; protein protein interaction; protein structure; regulatory mechanism; Saccharomyces cerevisiae; Trypanosoma cruzi; Adenylate Cyclase; Amino Acid Sequence; Animals; Base Sequence; Calcium; Catalytic Domain; Cloning, Molecular; Cyclic AMP; Dimerization; DNA Primers; Enzyme Activation; Molecular Sequence Data; Protein Binding; Protozoan Proteins; Reverse Transcriptase Polymerase Chain Reaction; Sequence Homology, Amino Acid; Signal Transduction; Trypanosoma cruzi; Bovinae; Mammalia; Protozoa; Saccharomyces; Saccharomyces cerevisiae; Trypanosoma; Trypanosoma cruzi
Año:2002
Volumen:277
Número:38
Página de inicio:35025
Página de fin:35034
DOI: http://dx.doi.org/10.1074/jbc.M204696200
Título revista:Journal of Biological Chemistry
Título revista abreviado:J. Biol. Chem.
ISSN:00219258
CODEN:JBCHA
CAS:Adenylate Cyclase, EC 4.6.1.1; Calcium, 7440-70-2; Cyclic AMP, 60-92-4; DNA Primers; Protozoan Proteins
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219258_v277_n38_p35025_DAngelo

Referencias:

  • Patel, T.B., Du, Z., Pierre, S., Cartin, L., Scholich, K., (2001) Gene (Amst.), 269, pp. 13-25
  • Hurley, J.H., (1999) J. Biol. Chem., 274, pp. 7599-7602
  • Lucas, K.A., Pitari, G.M., Kazerounian, S., Ruiz-Stewart, I., Park, J., Schulz, S., Kenneth, P., Waldman, S.A., (2000) Pharmacol. Rev., 52, pp. 375-414
  • Seebeck, T., Gong, K.W., Kunz, S., Schaub, R., Shalaby, T., Zoraghi, R., (2001) Int. J. Parasitol., 31, pp. 491-498
  • Naula, C., Seebeck, T., (2000) Parasitol. Today, 16, pp. 35-38
  • Taylor, M.C., Muhia, D.K., Baker, D.A., Mondragon, A., Schaap, P., Kelly, J.M., (1999) Mol. Biochem. Parasitol., 104, pp. 205-217
  • Rangel-Aldao, R., Allende, O., Triana, F., Piras, R., Henriquez, D., Piras, M., (1987) Mol. Biochem. Parasitol., 22, pp. 39-43
  • Flawiá, M.M., Tellez-Iñon, M.T., Torres, H.N., (1997) Parasitol. Today, 13, pp. 30-33
  • Fraidenraich, D., Peña, C., Isola, E.L., Lammel, E.M., Coso, O., Diaz Añel, A., Sandor, P., Flawiá, M.M., (1993) Proc. Natl. Acad. Sci. U. S. A., 90, pp. 10140-10144
  • Garcia, E.S., Gonzalez, M.S., De Azambuja, P., Baralle, F.E., Fraidenraich, D., Torres, H.N., Flawiá, M.M., (1995) Exp. Parasitol., 81, pp. 255-261
  • Ouaissi, A., Cornette, J., Schoneck, R., Plumas-Marty, B., Taibi, A., Loyens, M., Capron, A., (1992) Eur. J. Cell Biol., 59, pp. 68-79
  • Rangel-Aldao, R., Triana, F., Fernandez, V., Comach, G., Abate, T., Montoreano, R., (1988) Biochem. Int., 17, pp. 337-344
  • Rolin, S., Paindavoine, P., Hanocq-Quertier, J., Hanocq, F., Claes, Y., Le Ray, D., Overath, P., Pays, E., (1993) Mol. Biochem. Parasitol., 61, pp. 115-125
  • Vassella, E., Reuner, B., Yutzy, B., Boshart, M., (1997) J. Cell Sci., 110, pp. 2661-2671
  • Stojdl, D.F., Clarke, M.W., (1996) Exp. Parasitol., 83, pp. 134-146
  • Lammel, E.M., Barbieri, M.A., Wilkowsky, S.E., Bertini, F., Isola, E.L., (1996) Exp. Parasitol., 83, pp. 240-249
  • Engman, D.M., Krause, K.H., Blumin, J.H., Kim, K.S., Kirchhoff, L.V., Donelson, J.E., (1989) J. Biol. Chem., 264, pp. 18627-18631
  • Wu, Y., Haghighat, N.G., Ruben, L., (1992) Biochem. J., 287, pp. 187-193
  • Wu, Y., Deford, J., Benjamin, R., Lee, M.G., Ruben, L., (1994) Biochem. J., 304, pp. 833-841
  • Maldonado, R.A., Linss, J., Thomaz, N., Olson, C.L., Engman, D.M., Goldenberg, S., (1997) Exp. Parasitol., 86, pp. 200-205
  • Godsel, L.M., Engman, D.M., (1999) EMBO J., 18, pp. 2057-2065
  • Ridgley, E., Webster, P., Patton, C., Ruben, L., (2000) Mol. Biochem. Parasitol., 109, pp. 195-201
  • Pereira, C.A., Alonso, G.D., Paveto, M.C., Iribarren, A., Cabanas, M.L., Torres, H.N., Flawiá, M.M., (2000) J. Biol. Chem., 275, pp. 1495-1501
  • Zingales, B., Rondinelli, E., Degrave, W., Da Silevira, F., Levin, M., Le Paslier, D., Modabber, F., Frash, A.C., (1997) Parasitol. Today, 13, pp. 16-22
  • Gietz, R.D., Schiestl, R.H., (1991) Yeast, 7, pp. 253-263
  • Flawiá, M.M., Kornblihtt, A.R., Reig, J.A., Torruella, M., Torres, H.N., (1983) J. Biol. Chem., 258, pp. 8255-8259
  • Bartel, P., Chien, C., Sternglanz, R., Fields, S., (1993) Cellular Interactions in Development: A Practical Approach, pp. 153-179. , (Hartley, D. A., ed), Oxford University Press, Oxford, UK
  • Gomez, E.B., Santori, M.I., Laría, S., Engel, J.C., Swindle, J., Eisen, H., Szankasi, P., Téllez-Iñon, M.T., (2001) Mol. Biochem. Parasitol., 113, pp. 97-108
  • Pestka, S., Daugherty, B.L., Jung, V., Hotta, K., Pestka, R.K., (1984) Genetics, 81, pp. 7525-7528
  • Tesmer, J.J., Sprang, S.R., (1998) Curr. Opin. Struct. Biol., 8, pp. 713-719
  • Dessauer, C.W., Gilman, A.G., (1997) J. Biol. Chem., 272, pp. 27787-27795
  • Tesmer, J.J., Dessauer, C.W., Sunahara, R.K., Murray, L.D., Johnson, R.A., Gilman, A.G., Sprang, S.R., (2000) Biochemistry, 39, pp. 14464-14471
  • Désaubry, L., Shoshani, I., Johnson, R.A., (1996) J. Biol. Chem., 271, pp. 2380-2382
  • Johnson, R.A., Shoshani, I., (1990) J. Biol. Chem., 265, pp. 19035-19039
  • Torruella, M., Flawiá, M.M., Eisenschlos, C., Molina, Y., Vedia, L., Rubinstein, C.P., Torres, H.N., (1986) Biochem. J., 234, pp. 145-150
  • Eisenschlos, C., Flawiá, M.M., Torruella, M., Torres, H.N., (1986) Biochem. J., 236, pp. 185-191
  • Martin, R.B., Voorheis, P.H., Kennedy, E.L., (1978) Biochem. J., 175, pp. 207-212
  • Paindavoine, P., Rolin, S., Van Assel, S., Geuskens, M., Jauniaux, J.C., Dinsart, C., Huet, G., Pays, E., (1992) Mol. Cell. Biol., 12, pp. 1218-1225
  • Docampo, R., (1993) Biol. Res., 26, pp. 189-196
  • Vercesi, A.E., Hoffman, M.E., Bernades, C.F., Docampo, R., (1991) Cell Calcium, 12, pp. 361-369
  • Vanhamme, L., Pays, E., (1995) Microbiol. Rev., 59, pp. 223-240
  • Boucher, N., Wu, Y., Dumas, C., Dube, M., Sereno, D., Breton, M., Papadopoulou, B., (2002) J. Biol. Chem., 277, pp. 19511-19520
  • D'Orso, I., Frash, A.C., (2001) J. Biol. Chem., 276, pp. 15783-15793
  • Teixeira, S.M., Russell, D.G., Kirchhoff, L.V., Donelson, J.E., (1994) J. Biol. Chem., 269, pp. 20509-20516
  • Diaz Anel, A.M., Rossi, M.S., Espinosa, J.M., Guida, C., Freitas, F.A., Kornblihtt, A.R., Zingales, B., Torres, H.N., (2000) J. Eukaryot. Microbiol., 47, pp. 555-560
  • Sanchez, M.A., Zeoli, D., Klamos, E.M., Kavanaugh, M.P., Landfear, S.M., (1995) J. Biol. Chem., 270, pp. 17551-17558
  • Naula, C., Schaub, R., Leech, V., Melville, S., Seebeck, T., (2001) Mol. Biochem. Parasitol., 112, pp. 19-28
  • Landfear, S.M., Ignatushchenko, M., (2001) Mol. Biochem. Parasitol., 115, pp. 1-17
  • Bastin, P., MacRae, T.H., Francis, S.B., Matthews, K.R., Gull, K., (1999) Mol. Cell Biol., 19, pp. 8191-8200
  • Maga, J.A., LeBowitz, J.H., (1999) Trends Cell Biol., 9, pp. 409-413

Citas:

---------- APA ----------
D'Angelo, M.A., Montagna, A.E., Sanguineti, S., Torres, H.N. & Flawiá, M.M. (2002) . A novel calcium-stimulated adenylyl cyclase from Trypanosoma cruzi, which interacts with the structural flagellar protein paraflagellar rod. Journal of Biological Chemistry, 277(38), 35025-35034.
http://dx.doi.org/10.1074/jbc.M204696200
---------- CHICAGO ----------
D'Angelo, M.A., Montagna, A.E., Sanguineti, S., Torres, H.N., Flawiá, M.M. "A novel calcium-stimulated adenylyl cyclase from Trypanosoma cruzi, which interacts with the structural flagellar protein paraflagellar rod" . Journal of Biological Chemistry 277, no. 38 (2002) : 35025-35034.
http://dx.doi.org/10.1074/jbc.M204696200
---------- MLA ----------
D'Angelo, M.A., Montagna, A.E., Sanguineti, S., Torres, H.N., Flawiá, M.M. "A novel calcium-stimulated adenylyl cyclase from Trypanosoma cruzi, which interacts with the structural flagellar protein paraflagellar rod" . Journal of Biological Chemistry, vol. 277, no. 38, 2002, pp. 35025-35034.
http://dx.doi.org/10.1074/jbc.M204696200
---------- VANCOUVER ----------
D'Angelo, M.A., Montagna, A.E., Sanguineti, S., Torres, H.N., Flawiá, M.M. A novel calcium-stimulated adenylyl cyclase from Trypanosoma cruzi, which interacts with the structural flagellar protein paraflagellar rod. J. Biol. Chem. 2002;277(38):35025-35034.
http://dx.doi.org/10.1074/jbc.M204696200