Independent genes coding for three acidic proteins of the large ribosomal subunit from Saccharomyces cerevisiae.
The yeast ribosome contains three acidic proteins, L44, L44', and L45, closely related from a structural point of view, that seem to play a functional role similar to that of proteins L7 and L12 in the bacterial ribosome. By screening a cDNA bank in lambda gt11 with specific polyclonal and monoclonal antibodies, recombinant phages expressing each one of the acidic proteins have been cloned. A unique copy of each gene is detected using the phage cDNA inserts as probes in nitrocellulose blots of yeast DNA digested with different restriction enzymes. The inserts were subcloned in the plasmid pUC19, and their physical maps and nucleotide sequences were determined. By using the cDNA inserts as probes in genomic DNA banks, DNA fragments carrying the acidic protein genes have been cloned, characterized, and sequenced. The results conclusively show that the three yeast acidic proteins are coded by independent genes and are not the result of a post-translational modification of the product of a unique gene, as in bacteria. Like most ribosomal protein genes, the gene for protein L44' has an intron and two upstream stimulatory boxes (UASrpg) fitting closely to the consensus sequence. The genes coding for proteins L44 and L45 lack introns and seem also exceptional in other characteristics of their sequences. Proteins L44 and L45 have amino acid sequences with about 80% similarity. Protein L44' is only 63% similar to the other two polypeptides. The three proteins have highly conserved carboxyl termini comprising the last 30 amino acids, and the first 10 amino acids of L44 and L45 are identical. The results cast doubts about the possibility of a similar role for the different acidic ribosomal proteins.
Remacha
M
M
Centro de Biología Molecular, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
Sáenz-Robles
M T
MT
Vilella
M D
MD
Ballesta
J P
JP
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D004271:DNA, Fungal / Q000235:genetics; D005796:Genes; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D012269:Ribosomal Proteins / Q000235:genetics*; D012270:Ribosomes / Q000648:ultrastructure; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D004251:DNA Transposable Elements; D004271:DNA, Fungal; D012269:Ribosomal Proteins