RESUMEN

N(α)-Acetyltransferases (NATs) cause the N(α)-acetylation of the majority of eukaryotic proteins during their translation, although the functions of this modification have been largely unexplored. In yeast (Saccharomyces cerevisiae), four NATs have been identified: NatA, NatB, NatC, and NatD. In this study, the N(α)-acetylation status of ribosomal protein was analyzed using NAT mutants combined with two-dimensional difference gel electrophoresis (2D-DIGE) and mass spectrometry (MS). A total of 60 ribosomal proteins were identified, of which 17 were N(α)-acetylated by NatA, and two by NatB. The N(α)-acetylation of two of these, S17 and L23, by NatA was not previously observed. Furthermore, we tested the effect of ribosomal protein N(α)-acetylation on protein synthesis using the purified ribosomes from each NAT mutant. It was found that the protein synthesis activities of ribosomes from NatA and NatB mutants were decreased by 27% and 23%, respectively, as compared to that of the normal strain. Furthermore, we have shown that ribosomal protein N(α)-acetylation by NatA influences translational fidelity in the presence of paromomycin. These results suggest that ribosomal protein N(α)-acetylation is necessary to maintain the ribosome's protein synthesis function.

Asunto(s)

Acetiltransferasas/metabolismo , Proteínas Fúngicas/metabolismo , Biosíntesis de Proteínas , Procesamiento Proteico-Postraduccional/fisiología , Proteínas Ribosómicas/metabolismo , Acetilación , Acetiltransferasas/análisis , Acetiltransferasas/genética , Acetiltransferasas/fisiología , Secuencia de Aminoácidos , Secuencia de Bases , Proliferación Celular , Proteínas Fúngicas/análisis , Proteínas Fúngicas/genética , Modelos Moleculares , Datos de Secuencia Molecular , Organismos Modificados Genéticamente , Polirribosomas/genética , Polirribosomas/metabolismo , Biosíntesis de Proteínas/genética , Biosíntesis de Proteínas/fisiología , Procesamiento Proteico-Postraduccional/genética , Proteínas Ribosómicas/análisis , Proteínas Ribosómicas/genética , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/crecimiento & desarrollo , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Electroforesis Bidimensional Diferencial en Gel

Asunto(s)

Proteínas/química , Análisis de Secuencia de Proteína/métodos , Acetilación , Aminopeptidasas/química , Metanol/química , Péptido Hidrolasas/química , Pyrococcus/enzimología , Ácido Pirrolidona Carboxílico/química , Ácido Trifluoroacético/química