Trypanosoma cruzi TcSRPK, the first protozoan member of the SRPK family, is biochemically and functionally conserved with metazoan SR protein-specific kinases

A novel SR protein-specific kinase (SRPK) from the SRPK family was identified for the first time in a protozoan organism. The primary structure of the protein, named TcSRPK, presents a significant degree of identity with other metazoan members of the family. In vitro phosphorylation experiments show...

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Autores principales:	Portal, Daniel, Lobo, Guillermo Sebastián, Kadener, Sebastián, Espinosa, Joaquín Maximiliano, Pereira, Claudio Alejandro, Kornblihtt, Alberto Rodolfo, Flawiá, Mirtha María, Torres, Héctor Norberto
Publicado:	2003
Materias:	SR protein kinase SR-network SRPK Trans-splicing Trypanosoma cruzi cell extract epitope fibronectin isoenzyme monoclonal antibody monoclonal antibody 104 protein kinase protein S 100 protozoal protein ribonucleoprotein RNA serine arginine protein serine arginine protein kinase splicing factor 2 unclassified drug alternative RNA splicing article cell nucleus controlled study enzyme phosphorylation enzyme specificity exon fungal strain gene HeLa cell human human cell in vitro study in vivo study inhibition kinetics metazoon molecular recognition nonhuman nucleotide sequence phenotype priority journal protein expression protein family protein function protein structure Schizosaccharomyces pombe sequence homology Amino Acid Sequence Animals Antibodies, Monoclonal Cloning, Molecular Genes, Protozoan Globins Hela Cells Humans Molecular Sequence Data Protein Structure, Tertiary Protein-Serine-Threonine Kinases Schizosaccharomyces Sequence Alignment Sequence Homology, Amino Acid Species Specificity Metazoa Protozoa Trypanosoma
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v127_n1_p9_Portal http://hdl.handle.net/20.500.12110/paper_01666851_v127_n1_p9_Portal
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

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spelling	paper:paper_01666851_v127_n1_p9_Portal2023-06-08T15:15:54Z Trypanosoma cruzi TcSRPK, the first protozoan member of the SRPK family, is biochemically and functionally conserved with metazoan SR protein-specific kinases Portal, Daniel Lobo, Guillermo Sebastián Kadener, Sebastián Espinosa, Joaquín Maximiliano Pereira, Claudio Alejandro Kornblihtt, Alberto Rodolfo Flawiá, Mirtha María Torres, Héctor Norberto SR protein kinase SR-network SRPK Trans-splicing Trypanosoma cruzi cell extract epitope fibronectin isoenzyme monoclonal antibody monoclonal antibody 104 protein kinase protein S 100 protozoal protein ribonucleoprotein RNA serine arginine protein serine arginine protein kinase splicing factor 2 unclassified drug alternative RNA splicing article cell nucleus controlled study enzyme phosphorylation enzyme specificity exon fungal strain gene HeLa cell human human cell in vitro study in vivo study inhibition kinetics metazoon molecular recognition nonhuman nucleotide sequence phenotype priority journal protein expression protein family protein function protein structure Schizosaccharomyces pombe sequence homology Trypanosoma cruzi Amino Acid Sequence Animals Antibodies, Monoclonal Cloning, Molecular Genes, Protozoan Globins Hela Cells Humans Molecular Sequence Data Protein Structure, Tertiary Protein-Serine-Threonine Kinases Schizosaccharomyces Sequence Alignment Sequence Homology, Amino Acid Species Specificity Trypanosoma cruzi Metazoa Protozoa Schizosaccharomyces Schizosaccharomyces pombe Trypanosoma Trypanosoma cruzi A novel SR protein-specific kinase (SRPK) from the SRPK family was identified for the first time in a protozoan organism. The primary structure of the protein, named TcSRPK, presents a significant degree of identity with other metazoan members of the family. In vitro phosphorylation experiments showed that TcSRPK has the same substrate specificity relative to other SRPKs. TcSRPK was able to generate a mAb104-recognized phosphoepitope, a SRPK landmark. Expression of TcSRPK in different Schizosaccharomyces pombe strains lead to conserved phenotypes, indicating that TcSRPK is a functional homologue of metazoan SRPKs. In functional alternative splicing assays in vivo in HeLa cells, TcSRPK enhanced SR protein-dependent inclusion of the EDI exon of the fibronectin minigene. When tested in vitro, it inhibited splicing either on nuclear extracts or on splicing-deficient S100 extracts complemented with ASF/SF2. This inhibition was similar to that observed with human SRPK1. This work constitutes the first report of a member of this family of proteins and the existence of an SR-network in a protozoan organism. The implications in the origins and control of splicing are discussed. © 2002 Elsevier Science B.V. All rights reserved. Fil:Portal, D. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Lobo, G.S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Kadener, S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Espinosa, J.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Pereira, C.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Kornblihtt, A.R. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Flawiá, M.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Torres, H.N. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2003 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v127_n1_p9_Portal http://hdl.handle.net/20.500.12110/paper_01666851_v127_n1_p9_Portal
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	SR protein kinase SR-network SRPK Trans-splicing Trypanosoma cruzi cell extract epitope fibronectin isoenzyme monoclonal antibody monoclonal antibody 104 protein kinase protein S 100 protozoal protein ribonucleoprotein RNA serine arginine protein serine arginine protein kinase splicing factor 2 unclassified drug alternative RNA splicing article cell nucleus controlled study enzyme phosphorylation enzyme specificity exon fungal strain gene HeLa cell human human cell in vitro study in vivo study inhibition kinetics metazoon molecular recognition nonhuman nucleotide sequence phenotype priority journal protein expression protein family protein function protein structure Schizosaccharomyces pombe sequence homology Trypanosoma cruzi Amino Acid Sequence Animals Antibodies, Monoclonal Cloning, Molecular Genes, Protozoan Globins Hela Cells Humans Molecular Sequence Data Protein Structure, Tertiary Protein-Serine-Threonine Kinases Schizosaccharomyces Sequence Alignment Sequence Homology, Amino Acid Species Specificity Trypanosoma cruzi Metazoa Protozoa Schizosaccharomyces Schizosaccharomyces pombe Trypanosoma Trypanosoma cruzi
spellingShingle	SR protein kinase SR-network SRPK Trans-splicing Trypanosoma cruzi cell extract epitope fibronectin isoenzyme monoclonal antibody monoclonal antibody 104 protein kinase protein S 100 protozoal protein ribonucleoprotein RNA serine arginine protein serine arginine protein kinase splicing factor 2 unclassified drug alternative RNA splicing article cell nucleus controlled study enzyme phosphorylation enzyme specificity exon fungal strain gene HeLa cell human human cell in vitro study in vivo study inhibition kinetics metazoon molecular recognition nonhuman nucleotide sequence phenotype priority journal protein expression protein family protein function protein structure Schizosaccharomyces pombe sequence homology Trypanosoma cruzi Amino Acid Sequence Animals Antibodies, Monoclonal Cloning, Molecular Genes, Protozoan Globins Hela Cells Humans Molecular Sequence Data Protein Structure, Tertiary Protein-Serine-Threonine Kinases Schizosaccharomyces Sequence Alignment Sequence Homology, Amino Acid Species Specificity Trypanosoma cruzi Metazoa Protozoa Schizosaccharomyces Schizosaccharomyces pombe Trypanosoma Trypanosoma cruzi Portal, Daniel Lobo, Guillermo Sebastián Kadener, Sebastián Espinosa, Joaquín Maximiliano Pereira, Claudio Alejandro Kornblihtt, Alberto Rodolfo Flawiá, Mirtha María Torres, Héctor Norberto Trypanosoma cruzi TcSRPK, the first protozoan member of the SRPK family, is biochemically and functionally conserved with metazoan SR protein-specific kinases
topic_facet	SR protein kinase SR-network SRPK Trans-splicing Trypanosoma cruzi cell extract epitope fibronectin isoenzyme monoclonal antibody monoclonal antibody 104 protein kinase protein S 100 protozoal protein ribonucleoprotein RNA serine arginine protein serine arginine protein kinase splicing factor 2 unclassified drug alternative RNA splicing article cell nucleus controlled study enzyme phosphorylation enzyme specificity exon fungal strain gene HeLa cell human human cell in vitro study in vivo study inhibition kinetics metazoon molecular recognition nonhuman nucleotide sequence phenotype priority journal protein expression protein family protein function protein structure Schizosaccharomyces pombe sequence homology Trypanosoma cruzi Amino Acid Sequence Animals Antibodies, Monoclonal Cloning, Molecular Genes, Protozoan Globins Hela Cells Humans Molecular Sequence Data Protein Structure, Tertiary Protein-Serine-Threonine Kinases Schizosaccharomyces Sequence Alignment Sequence Homology, Amino Acid Species Specificity Trypanosoma cruzi Metazoa Protozoa Schizosaccharomyces Schizosaccharomyces pombe Trypanosoma Trypanosoma cruzi
description	A novel SR protein-specific kinase (SRPK) from the SRPK family was identified for the first time in a protozoan organism. The primary structure of the protein, named TcSRPK, presents a significant degree of identity with other metazoan members of the family. In vitro phosphorylation experiments showed that TcSRPK has the same substrate specificity relative to other SRPKs. TcSRPK was able to generate a mAb104-recognized phosphoepitope, a SRPK landmark. Expression of TcSRPK in different Schizosaccharomyces pombe strains lead to conserved phenotypes, indicating that TcSRPK is a functional homologue of metazoan SRPKs. In functional alternative splicing assays in vivo in HeLa cells, TcSRPK enhanced SR protein-dependent inclusion of the EDI exon of the fibronectin minigene. When tested in vitro, it inhibited splicing either on nuclear extracts or on splicing-deficient S100 extracts complemented with ASF/SF2. This inhibition was similar to that observed with human SRPK1. This work constitutes the first report of a member of this family of proteins and the existence of an SR-network in a protozoan organism. The implications in the origins and control of splicing are discussed. © 2002 Elsevier Science B.V. All rights reserved.
author	Portal, Daniel Lobo, Guillermo Sebastián Kadener, Sebastián Espinosa, Joaquín Maximiliano Pereira, Claudio Alejandro Kornblihtt, Alberto Rodolfo Flawiá, Mirtha María Torres, Héctor Norberto
author_facet	Portal, Daniel Lobo, Guillermo Sebastián Kadener, Sebastián Espinosa, Joaquín Maximiliano Pereira, Claudio Alejandro Kornblihtt, Alberto Rodolfo Flawiá, Mirtha María Torres, Héctor Norberto
author_sort	Portal, Daniel
title	Trypanosoma cruzi TcSRPK, the first protozoan member of the SRPK family, is biochemically and functionally conserved with metazoan SR protein-specific kinases
title_short	Trypanosoma cruzi TcSRPK, the first protozoan member of the SRPK family, is biochemically and functionally conserved with metazoan SR protein-specific kinases
title_full	Trypanosoma cruzi TcSRPK, the first protozoan member of the SRPK family, is biochemically and functionally conserved with metazoan SR protein-specific kinases
title_fullStr	Trypanosoma cruzi TcSRPK, the first protozoan member of the SRPK family, is biochemically and functionally conserved with metazoan SR protein-specific kinases
title_full_unstemmed	Trypanosoma cruzi TcSRPK, the first protozoan member of the SRPK family, is biochemically and functionally conserved with metazoan SR protein-specific kinases
title_sort	trypanosoma cruzi tcsrpk, the first protozoan member of the srpk family, is biochemically and functionally conserved with metazoan sr protein-specific kinases
publishDate	2003
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v127_n1_p9_Portal http://hdl.handle.net/20.500.12110/paper_01666851_v127_n1_p9_Portal
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Trypanosoma cruzi TcSRPK, the first protozoan member of the SRPK family, is biochemically and functionally conserved with metazoan SR protein-specific kinases

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