Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1
Early in the assembly of the spliceosome of eukaryotes the branch-point binding protein (BBP, also called SF1) recognizes the branch point sequence, whereas the heterodimer U2AF consisting of a 65 and a 35 kDa subunit, contacts the polypyrimidine tract and the AG splice site, respectively. Herein, w...
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paper:paper_01666851_v164_n2_p137_Vazquez2023-06-08T15:16:00Z Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1 Vazquez, Martín Pablo Bercovich, Natalia Trans-splicing Trypanosoma brucei Trypanosoma cruzi U2 auxiliary factor protein Sf1 protein u2af35 protein U2AF65 protozoal protein unclassified drug article cell fractionation controlled study molecular cloning nonhuman priority journal protein analysis protein expression protein function protein localization protein protein interaction RNA splicing Trypanosoma brucei Trypanosoma cruzi Amino Acid Sequence Animals Cell Fractionation Cloning, Molecular Conserved Sequence Gene Expression Gene Silencing Models, Molecular Molecular Sequence Data Protein Interaction Mapping Protein Structure, Tertiary Protozoan Proteins RNA Splicing RNA-Binding Proteins Sequence Homology, Amino Acid Trypanosoma brucei brucei Trypanosoma cruzi Two-Hybrid System Techniques Eukaryota Mammalia Trypanosoma brucei Trypanosoma cruzi Trypanosomatidae Early in the assembly of the spliceosome of eukaryotes the branch-point binding protein (BBP, also called SF1) recognizes the branch point sequence, whereas the heterodimer U2AF consisting of a 65 and a 35 kDa subunit, contacts the polypyrimidine tract and the AG splice site, respectively. Herein, we identified, cloned and expressed the Trypanosoma cruzi and Trypanosoma brucei U2AF35, U2AF65 and SF1. Trypanosomatid U2AF65 strongly diverged from yeast and human homologues. On the contrary, trypanosomatid SF1 was conserved but lacked the C-terminal sequence present in the mammalian protein. Yeast two hybrid approaches were used to assess their interactions. The interaction between U2AF35 and U2AF65 was very weak or not detectable. However, as in other eukaryotes, the interaction between U2AF65 and SF1 was strong. At the cellular level, these results were confirmed by fractionation and affinity-selection experiments in which SF1 and U2AF65 co-fractionated in a complex of approximately 400 kDa and U2AF65 was affinity-selected with TAP tagged SF1, but not with TAP tagged U2AF35. Silencing of the three factors affected growth and trans-splicing in the first step of this reaction. Trypanosomes are the first described example of eukaryotic cells in which the interaction of two expressed U2AF factors seemed to be very weak, or even undetectable. © 2009 Elsevier B.V. All rights reserved. Fil:Vazquez, M.P. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Bercovich, N. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2009 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v164_n2_p137_Vazquez http://hdl.handle.net/20.500.12110/paper_01666851_v164_n2_p137_Vazquez |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Trans-splicing Trypanosoma brucei Trypanosoma cruzi U2 auxiliary factor protein Sf1 protein u2af35 protein U2AF65 protozoal protein unclassified drug article cell fractionation controlled study molecular cloning nonhuman priority journal protein analysis protein expression protein function protein localization protein protein interaction RNA splicing Trypanosoma brucei Trypanosoma cruzi Amino Acid Sequence Animals Cell Fractionation Cloning, Molecular Conserved Sequence Gene Expression Gene Silencing Models, Molecular Molecular Sequence Data Protein Interaction Mapping Protein Structure, Tertiary Protozoan Proteins RNA Splicing RNA-Binding Proteins Sequence Homology, Amino Acid Trypanosoma brucei brucei Trypanosoma cruzi Two-Hybrid System Techniques Eukaryota Mammalia Trypanosoma brucei Trypanosoma cruzi Trypanosomatidae |
| spellingShingle |
Trans-splicing Trypanosoma brucei Trypanosoma cruzi U2 auxiliary factor protein Sf1 protein u2af35 protein U2AF65 protozoal protein unclassified drug article cell fractionation controlled study molecular cloning nonhuman priority journal protein analysis protein expression protein function protein localization protein protein interaction RNA splicing Trypanosoma brucei Trypanosoma cruzi Amino Acid Sequence Animals Cell Fractionation Cloning, Molecular Conserved Sequence Gene Expression Gene Silencing Models, Molecular Molecular Sequence Data Protein Interaction Mapping Protein Structure, Tertiary Protozoan Proteins RNA Splicing RNA-Binding Proteins Sequence Homology, Amino Acid Trypanosoma brucei brucei Trypanosoma cruzi Two-Hybrid System Techniques Eukaryota Mammalia Trypanosoma brucei Trypanosoma cruzi Trypanosomatidae Vazquez, Martín Pablo Bercovich, Natalia Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1 |
| topic_facet |
Trans-splicing Trypanosoma brucei Trypanosoma cruzi U2 auxiliary factor protein Sf1 protein u2af35 protein U2AF65 protozoal protein unclassified drug article cell fractionation controlled study molecular cloning nonhuman priority journal protein analysis protein expression protein function protein localization protein protein interaction RNA splicing Trypanosoma brucei Trypanosoma cruzi Amino Acid Sequence Animals Cell Fractionation Cloning, Molecular Conserved Sequence Gene Expression Gene Silencing Models, Molecular Molecular Sequence Data Protein Interaction Mapping Protein Structure, Tertiary Protozoan Proteins RNA Splicing RNA-Binding Proteins Sequence Homology, Amino Acid Trypanosoma brucei brucei Trypanosoma cruzi Two-Hybrid System Techniques Eukaryota Mammalia Trypanosoma brucei Trypanosoma cruzi Trypanosomatidae |
| description |
Early in the assembly of the spliceosome of eukaryotes the branch-point binding protein (BBP, also called SF1) recognizes the branch point sequence, whereas the heterodimer U2AF consisting of a 65 and a 35 kDa subunit, contacts the polypyrimidine tract and the AG splice site, respectively. Herein, we identified, cloned and expressed the Trypanosoma cruzi and Trypanosoma brucei U2AF35, U2AF65 and SF1. Trypanosomatid U2AF65 strongly diverged from yeast and human homologues. On the contrary, trypanosomatid SF1 was conserved but lacked the C-terminal sequence present in the mammalian protein. Yeast two hybrid approaches were used to assess their interactions. The interaction between U2AF35 and U2AF65 was very weak or not detectable. However, as in other eukaryotes, the interaction between U2AF65 and SF1 was strong. At the cellular level, these results were confirmed by fractionation and affinity-selection experiments in which SF1 and U2AF65 co-fractionated in a complex of approximately 400 kDa and U2AF65 was affinity-selected with TAP tagged SF1, but not with TAP tagged U2AF35. Silencing of the three factors affected growth and trans-splicing in the first step of this reaction. Trypanosomes are the first described example of eukaryotic cells in which the interaction of two expressed U2AF factors seemed to be very weak, or even undetectable. © 2009 Elsevier B.V. All rights reserved. |
| author |
Vazquez, Martín Pablo Bercovich, Natalia |
| author_facet |
Vazquez, Martín Pablo Bercovich, Natalia |
| author_sort |
Vazquez, Martín Pablo |
| title |
Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1 |
| title_short |
Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1 |
| title_full |
Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1 |
| title_fullStr |
Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1 |
| title_full_unstemmed |
Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1 |
| title_sort |
functional characterization and protein-protein interactions of trypanosome splicing factors u2af35, u2af65 and sf1 |
| publishDate |
2009 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v164_n2_p137_Vazquez http://hdl.handle.net/20.500.12110/paper_01666851_v164_n2_p137_Vazquez |
| work_keys_str_mv |
AT vazquezmartinpablo functionalcharacterizationandproteinproteininteractionsoftrypanosomesplicingfactorsu2af35u2af65andsf1 AT bercovichnatalia functionalcharacterizationandproteinproteininteractionsoftrypanosomesplicingfactorsu2af35u2af65andsf1 |
| _version_ |
1768545324205342720 |