In vivo analysis of the acidic ribosomal proteins BmP1 and BmP2 of the silkworm Bombyx mori in the yeast Saccharomyces cerevisiae.
In the silkworm Bombyx mori the ribosomal stalk P-protein family consists of two low MW acidic proteins, BmP1 and BmP2, and of one higher MW protein, BmP0, as shown by electrophoretical and immunoblotting western blot analysis of purified ribosomes. Treatment of ribosomes with alkaline phosphatase followed by electrofocusing shifted the isoelectric points to higher pH, implying phosphorylation of the proteins. The cDNAs encoding BmP1 and BmP2 proteins were constructed and expressed in the Saccharomyces cerevisiae mutant strains defective in either the endogenous P1 or P2 proteins. The recombinant silkworm proteins could complement the absence of the homologous yeast proteins and were incorporated to the ribosomes of the transformed strains, helping the binding of the remaining endogenous acidic proteins, present in the cytoplasm in different extent. Thus, BmP1 was able to replace YP1alpha, preferentially binding YP2beta to the ribosome, while BmP2 replaced both yeast P2 proteins and induced the binding of both YP1alpha and YP1beta.
Koumarianou
Petrina
P
University of Athens, Faculty of Biology, Department of Genetics and Biotechnology, Panepistimiopolis, 15701 Athens, Greece.
Marcos
Alberto Garcia
AG
Ballesta
Juan P G
JP
Kouyanou-Koutsoukou
Sophia
S
D000595:Amino Acid Sequence; D000818:Animals; D012831:Bombyx / Q000235:genetics / Q000378:metabolism*; D005260:Female; D005816:Genetic Complementation Test; D015151:Immunoblotting; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D006080:Ovarian Follicle / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000032:analysis / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000235:genetics / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D014170:Transformation, Genetic
D019476:Insect Proteins; D011994:Recombinant Proteins; D012269:Ribosomal Proteins