Characterization of interaction sites in the Saccharomyces cerevisiae ribosomal stalk components
The interactions among the yeast stalk components(P0, P1⍺, P1ꞵ, P2⍺ and P2ꞵ) and with EF-2 have been explored using immunoprecipitation, affinity chromatography and the two-hybrid system. No stable association was detected between acidic proteins of the same type. In contrast, P1⍺ and P1ꞵ were found...
| Autores: | , , , |
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| Tipo de recurso: | artículo |
| Estado: | Versión aceptada para publicación |
| Fecha de publicación: | 2002 |
| País: | España |
| Institución: | Universidad de Jaén |
| Repositorio: | RUJA. Repositorio Institucional de la Producción Científica de la Universidad de Jaén |
| OAI Identifier: | oai:ruja.ujaen.es:10953/3669 |
| Acceso en línea: | https://doi.org/10.1046/j.1365-2958.2002.03179.x https://onlinelibrary.wiley.com/doi/full/10.1046/j.1365-2958.2002.03179.x?sid=nlm%3A https://hdl.handle.net/10953/3669 |
| Access Level: | acceso abierto |
| Palabra clave: | Ribosomes Ribosomal stalk Phosphoproteins Translation Saccharomyces cerevisiae in vitro reconstitution |
| Sumario: | The interactions among the yeast stalk components(P0, P1⍺, P1ꞵ, P2⍺ and P2ꞵ) and with EF-2 have been explored using immunoprecipitation, affinity chromatography and the two-hybrid system. No stable association was detected between acidic proteins of the same type. In contrast, P1⍺ and P1ꞵ were found to interact with P2ꞵ and P2⍺ respectively. An interaction of P0 with P1 proteins, but not with P2 proteins, was also detected. This interaction is strongly increased with the P0 carboxyl end, which is able to form a pentameric complex with the four acidic proteins. The P1/P2 binding site has been located between residues 212 and 262 using different C-terminal P0 fragments. Immunoprecipitation shows the association of EF-2 with protein P0. However, the interaction is stronger with the P1/P2 proteins than with P0 in the two-hybrid assay. This interaction improves using the 100-amino-acid-long C-end of P0 and is even higher with the last 50 amino acids. The data indicate a specific association of P1⍺ with P2ꞵ and of P1ꞵ with P2⍺ rather than the dimerization of the acidic proteins found in prokaryotes. In addition, they suggest that stalk assembly begins by the interaction of the P1 proteins with P0. Moreover, as functional interactions of the complete P0 were found to increase using protein fragments, the data suggest that some active sites are exposed in the ribosome as a result of conformational changes that take place during stalk assembly and function. |
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