Phosphorylation controls binding of acidic proteins to the ribosome.
The replacement of each one of the eight serine residues present in the amino acid sequence of the Saccharomyces cerevisiae acidic ribosomal phosphoprotein YP2 beta (L45) by different amino acids has been performed by heteroduplex site-directed mutagenesis in the cloned gene. The mutated DNA was used to transform a yeast strain previously deprived of the original protein YP2 beta (L45) by gene disruption. The replacement of serine in position 19 by either alanine, aspartic acid, or threonine prevents in vivo phosphorylation of the protein and its interaction with the ribosome. The serine-19 mutated gene is unable to rescue the negative effect on the growth rate caused by elimination of the original protein in YP2 beta (L45) gene disrupted strains. The mutation of any one of the other seven serine residues has no effect on either the phosphorylation or the ribosome binding capacity of the protein, although replacement of serine-72 seems to increase the sensitivity of the polypeptide to degradation. These results provide strong evidence indicating that ribosomal protein phosphorylation plays an important part in the activity of the particle and that it supports the existence of a control mechanism of protein synthesis, which would regulate the level of phosphorylation of acidic proteins.
Naranda
T
T
Centro de Biologia Molecular, Consejo Superior de Investigaciones Científicas, Canto Blanco, Madrid, Spain.
Ballesta
J P
JP
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005800:Genes, Fungal; D007525:Isoelectric Focusing; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016297:Mutagenesis, Site-Directed; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D010768:Phosphoserine / Q000032:analysis; D015183:Restriction Mapping; D000090822:Ribosomal Protein L3; D012269:Ribosomal Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development; D014170:Transformation, Genetic
D010750:Phosphoproteins; D000090822:Ribosomal Protein L3; D012269:Ribosomal Proteins; D010768:Phosphoserine