Assembly of 60S ribosomal subunits is perturbed in temperature-sensitive yeast mutants defective in ribosomal protein L16.
Temperature-sensitive mutants defective in 60S ribosomal subunit protein L16 of Saccharomyces cerevisiae were isolated through hydroxylamine mutagenesis of the RPL16B gene and plasmid shuffling. Two heat-sensitive and two cold-sensitive isolates were characterized. The growth of the four mutants is inhibited at their restrictive temperatures. However, many of the cells remain viable if returned to their permissive temperatures. All of the mutants are deficient in 60S ribosomal subunits and therefore accumulate translational preinitiation complexes. Three of the mutants exhibit a shortage of mature 25S rRNA, and one accumulates rRNA precursors. The accumulation of rRNA precursors suggests that ribosome assembly may be slowed in this mutant. These phenotypes lead us to propose that mutants containing the rpl16b alleles are defective for 60S subunit assembly rather than function. In the mutant carrying the rpl16b-1 allele, ribosomes initiate translation at the noncanonical codon AUA, at least on the rpl16b-1 mRNA, bringing to light a possible connection between the rate and the fidelity of translation initiation.
Moritz
M
M
Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213.
Pulaski
B A
BA
Woolford
J L
JL
D000595:Amino Acid Sequence; D000818:Animals; D003062:Codon; D005656:Fungal Proteins / Q000032:analysis; D005800:Genes, Fungal; D005838:Genotype; D019811:Hydroxylamine; D006898:Hydroxylamines / Q000494:pharmacology; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011132:Polyribosomes / Q000378:metabolism; D012322:RNA Precursors / Q000235:genetics / Q000378:metabolism; D012335:RNA, Ribosomal / Q000235:genetics; D012269:Ribosomal Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D013696:Temperature
D003062:Codon; D005656:Fungal Proteins; D006898:Hydroxylamines; D012322:RNA Precursors; D012335:RNA, Ribosomal; D012269:Ribosomal Proteins; C027937:ribosomal protein L16; D019811:Hydroxylamine