An intragenic suppressor of cold sensitivity identifies potentially interacting bases in the peptidyl transferase center of Tetrahymena rRNA.
Peptidyl transfer of a growing peptide on a ribosome-bound transfer RNA (tRNA) to an incoming amino acyl tRNA is the central step in translation, and it may be catalyzed primarily by the large subunit (LSU) ribosomal RNA (rRNA). Genetic and biochemical evidence suggests that the central loop of domain V of the LSU rRNA plays a direct role in peptidyl transfer. It was previously found that a single base change at a universally conserved site in this region of the Tetrahymena thermophila LSU rRNA confers anisomycin resistance (an-r) as well as extremely slow growth, cold sensitivity, and aberrant cell morphology. Because anisomycin specifically inhibits peptidyl transfer, possibly by interfering with tRNA binding, it is likely that this mutant rRNA is defective in efficiently completing one of these steps. In the present study, we have isolated an intragenic suppressor mutation located only three bases away from the original mutation that partially reverses the slow growth and cold-sensitive phenotypes. These data imply that the functional interaction of these two bases is necessary for normal rRNA function, perhaps for peptidyl transfer or tRNA binding. These data provide the first demonstration of a functional interaction between bases within this rRNA region.
Sweeney
R
R
Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA.
Yao
M C
MC
D000818:Animals; D001482:Base Composition; D001483:Base Sequence; D002874:Chromosome Mapping; D003080:Cold Temperature; D016153:Genes, Suppressor / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D010458:Peptidyl Transferases / Q000235:genetics*; D010641:Phenotype; D016053:RNA, Protozoan / Q000235:genetics*; D012335:RNA, Ribosomal / Q000235:genetics*; D017422:Sequence Analysis, DNA; D016808:Tetrahymena thermophila / Q000235:genetics*; D014170:Transformation, Genetic
D016053:RNA, Protozoan; D012335:RNA, Ribosomal; D010458:Peptidyl Transferases