Calcineurin B of the human protozoan parasite Trypanosoma cruzi is involved in cell invasion

Jorge E. Araya, Alberto Cornejo, Patricio R. Orrego, Esteban M. Cordero, Mauro Cortéz, Héctor Olivares, Ivan Neira, Hernán Sagua, José Franco da Silveira, Nobuko Yoshida, Jorge González

Resultado de la investigación: Article

25 Citas (Scopus)

Resumen

During Trypanosoma cruzi cell invasion, signal transduction pathways are triggered in parasite and host cells, leading to a rise in intracellular Ca 2+ concentration. We posed the question whether calcineurin (CaN), in particular the functional regulatory subunit CaNB, a Ca 2+-binding EF-hand protein, was expressed in T. cruzi and whether it played a role in cell invasion. Here we report the cloning and characterization of CL strain CaNB gene, as well as the participation of CaNB in cell invasion. Treatment of metacyclic trypomastigotes (MT) or tissue-culture trypomastigotes (TCT) with the CaN inhibitors cyclosporin or cypermethrin strongly inhibited (62-64%) their entry into HeLa cells. In assays using anti-phospho-serine/threonine antibodies, a few proteins of MT were found to be dephosphorylated in a manner inhibitable by cyclosporin upon exposure to HeLa cell extract. The phosphatase activity of CaN was detected by a biochemical approach in both MT and TCT. Treatment of parasites with antisense phosphorothioate oligonucleotides directed to TcCaNB-CL, which reduced the expression of TcCaNB and affected TcCaN activity, resulted in ∼50% inhibition of HeLa cell entry by MT or TCT. Given that TcCaNB-CL may play a key role in cell invasion and differs considerably in its primary structure from the human CaNB, it might be considered as a potential chemotherapeutic target.

Idioma originalEnglish
Páginas (desde-hasta)892-900
Número de páginas9
PublicaciónMicrobes and Infection
Volumen10
N.º8
DOI
EstadoPublished - 1 jul 2008

Huella dactilar

Calcineurin
Trypanosoma cruzi
Parasites
HeLa Cells
Cyclosporine
Phosphorothioate Oligonucleotides
EF Hand Motifs
Antisense Oligonucleotides
Threonine
Cell Extracts
Serine
Organism Cloning
Signal Transduction
Proteins
Antibodies
Therapeutics
Genes

ASJC Scopus subject areas

  • Microbiology
  • Infectious Diseases

Citar esto

Araya, J. E., Cornejo, A., Orrego, P. R., Cordero, E. M., Cortéz, M., Olivares, H., ... González, J. (2008). Calcineurin B of the human protozoan parasite Trypanosoma cruzi is involved in cell invasion. Microbes and Infection, 10(8), 892-900. https://doi.org/10.1016/j.micinf.2008.05.003
Araya, Jorge E. ; Cornejo, Alberto ; Orrego, Patricio R. ; Cordero, Esteban M. ; Cortéz, Mauro ; Olivares, Héctor ; Neira, Ivan ; Sagua, Hernán ; Franco da Silveira, José ; Yoshida, Nobuko ; González, Jorge. / Calcineurin B of the human protozoan parasite Trypanosoma cruzi is involved in cell invasion. En: Microbes and Infection. 2008 ; Vol. 10, N.º 8. pp. 892-900.
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abstract = "During Trypanosoma cruzi cell invasion, signal transduction pathways are triggered in parasite and host cells, leading to a rise in intracellular Ca 2+ concentration. We posed the question whether calcineurin (CaN), in particular the functional regulatory subunit CaNB, a Ca 2+-binding EF-hand protein, was expressed in T. cruzi and whether it played a role in cell invasion. Here we report the cloning and characterization of CL strain CaNB gene, as well as the participation of CaNB in cell invasion. Treatment of metacyclic trypomastigotes (MT) or tissue-culture trypomastigotes (TCT) with the CaN inhibitors cyclosporin or cypermethrin strongly inhibited (62-64{\%}) their entry into HeLa cells. In assays using anti-phospho-serine/threonine antibodies, a few proteins of MT were found to be dephosphorylated in a manner inhibitable by cyclosporin upon exposure to HeLa cell extract. The phosphatase activity of CaN was detected by a biochemical approach in both MT and TCT. Treatment of parasites with antisense phosphorothioate oligonucleotides directed to TcCaNB-CL, which reduced the expression of TcCaNB and affected TcCaN activity, resulted in ∼50{\%} inhibition of HeLa cell entry by MT or TCT. Given that TcCaNB-CL may play a key role in cell invasion and differs considerably in its primary structure from the human CaNB, it might be considered as a potential chemotherapeutic target.",
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Araya, JE, Cornejo, A, Orrego, PR, Cordero, EM, Cortéz, M, Olivares, H, Neira, I, Sagua, H, Franco da Silveira, J, Yoshida, N & González, J 2008, 'Calcineurin B of the human protozoan parasite Trypanosoma cruzi is involved in cell invasion', Microbes and Infection, vol. 10, n.º 8, pp. 892-900. https://doi.org/10.1016/j.micinf.2008.05.003

Calcineurin B of the human protozoan parasite Trypanosoma cruzi is involved in cell invasion. / Araya, Jorge E.; Cornejo, Alberto; Orrego, Patricio R.; Cordero, Esteban M.; Cortéz, Mauro; Olivares, Héctor; Neira, Ivan; Sagua, Hernán; Franco da Silveira, José; Yoshida, Nobuko; González, Jorge.

En: Microbes and Infection, Vol. 10, N.º 8, 01.07.2008, p. 892-900.

Resultado de la investigación: Article

TY - JOUR

T1 - Calcineurin B of the human protozoan parasite Trypanosoma cruzi is involved in cell invasion

AU - Araya, Jorge E.

AU - Cornejo, Alberto

AU - Orrego, Patricio R.

AU - Cordero, Esteban M.

AU - Cortéz, Mauro

AU - Olivares, Héctor

AU - Neira, Ivan

AU - Sagua, Hernán

AU - Franco da Silveira, José

AU - Yoshida, Nobuko

AU - González, Jorge

PY - 2008/7/1

Y1 - 2008/7/1

N2 - During Trypanosoma cruzi cell invasion, signal transduction pathways are triggered in parasite and host cells, leading to a rise in intracellular Ca 2+ concentration. We posed the question whether calcineurin (CaN), in particular the functional regulatory subunit CaNB, a Ca 2+-binding EF-hand protein, was expressed in T. cruzi and whether it played a role in cell invasion. Here we report the cloning and characterization of CL strain CaNB gene, as well as the participation of CaNB in cell invasion. Treatment of metacyclic trypomastigotes (MT) or tissue-culture trypomastigotes (TCT) with the CaN inhibitors cyclosporin or cypermethrin strongly inhibited (62-64%) their entry into HeLa cells. In assays using anti-phospho-serine/threonine antibodies, a few proteins of MT were found to be dephosphorylated in a manner inhibitable by cyclosporin upon exposure to HeLa cell extract. The phosphatase activity of CaN was detected by a biochemical approach in both MT and TCT. Treatment of parasites with antisense phosphorothioate oligonucleotides directed to TcCaNB-CL, which reduced the expression of TcCaNB and affected TcCaN activity, resulted in ∼50% inhibition of HeLa cell entry by MT or TCT. Given that TcCaNB-CL may play a key role in cell invasion and differs considerably in its primary structure from the human CaNB, it might be considered as a potential chemotherapeutic target.

AB - During Trypanosoma cruzi cell invasion, signal transduction pathways are triggered in parasite and host cells, leading to a rise in intracellular Ca 2+ concentration. We posed the question whether calcineurin (CaN), in particular the functional regulatory subunit CaNB, a Ca 2+-binding EF-hand protein, was expressed in T. cruzi and whether it played a role in cell invasion. Here we report the cloning and characterization of CL strain CaNB gene, as well as the participation of CaNB in cell invasion. Treatment of metacyclic trypomastigotes (MT) or tissue-culture trypomastigotes (TCT) with the CaN inhibitors cyclosporin or cypermethrin strongly inhibited (62-64%) their entry into HeLa cells. In assays using anti-phospho-serine/threonine antibodies, a few proteins of MT were found to be dephosphorylated in a manner inhibitable by cyclosporin upon exposure to HeLa cell extract. The phosphatase activity of CaN was detected by a biochemical approach in both MT and TCT. Treatment of parasites with antisense phosphorothioate oligonucleotides directed to TcCaNB-CL, which reduced the expression of TcCaNB and affected TcCaN activity, resulted in ∼50% inhibition of HeLa cell entry by MT or TCT. Given that TcCaNB-CL may play a key role in cell invasion and differs considerably in its primary structure from the human CaNB, it might be considered as a potential chemotherapeutic target.

KW - Antisense oligonucleotides

KW - Ca mobilization

KW - Calcineurin B

KW - Cell invasion

KW - Inhibitors

KW - Phosphatase activity

KW - Trypanosoma cruzi

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U2 - 10.1016/j.micinf.2008.05.003

DO - 10.1016/j.micinf.2008.05.003

M3 - Article

VL - 10

SP - 892

EP - 900

JO - Microbes and Infection

JF - Microbes and Infection

SN - 1286-4579

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