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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todo:paper_01666851_v127_n1_p9_Portal2023-10-03T15:04:08Z Trypanosoma cruzi TcSRPK, the first protozoan member of the SRPK family, is biochemically and functionally conserved with metazoan SR protein-specific kinases Portal, D. Lobo, G.S. Kadener, S. Prasad, J. Espinosa, J.M. Pereira, C.A. Tang, Z. Lin, R.-J. Manley, J.L. Kornblihtt, A.R. Flawiá, M.M. Torres, H.N. 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. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar 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, D. Lobo, G.S. Kadener, S. Prasad, J. Espinosa, J.M. Pereira, C.A. Tang, Z. Lin, R.-J. Manley, J.L. Kornblihtt, A.R. Flawiá, M.M. Torres, H.N. 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. |
| format |
JOUR |
| author |
Portal, D. Lobo, G.S. Kadener, S. Prasad, J. Espinosa, J.M. Pereira, C.A. Tang, Z. Lin, R.-J. Manley, J.L. Kornblihtt, A.R. Flawiá, M.M. Torres, H.N. |
| author_facet |
Portal, D. Lobo, G.S. Kadener, S. Prasad, J. Espinosa, J.M. Pereira, C.A. Tang, Z. Lin, R.-J. Manley, J.L. Kornblihtt, A.R. Flawiá, M.M. Torres, H.N. |
| author_sort |
Portal, D. |
| 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 |
| url |
http://hdl.handle.net/20.500.12110/paper_01666851_v127_n1_p9_Portal |
| work_keys_str_mv |
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