Dinman J D, Wickner R B
Section on Genetics of Simple Eukaryotes, National Institute of Diabetes, Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA.
Genetics. 1995 Sep;141(1):95-105. doi: 10.1093/genetics/141.1.95.
Chromosomal mutants (maintenance of frame = mof) in which the efficiency of -1 ribosomal frameshifting is increased can be isolated using constructs in which lacZ expression is dependent upon a -1 shift of reading frame. We isolate a new mof mutation, mof9, in Saccharomyces cerevisiae and show that it is complemented by both single and multi-copy 5 S rDNA clones. Two independent insertion mutations in the rDNA locus (rDNA::LEU2 and rDNA::URA3) also display the Mof- phenotype and are also complemented by single and multi-copy 5 S rDNA clones. Mutant 5 S rRNAs expressed from a plasmid as 20-50% of total 5 S rRNA in a wild-type host also induced the Mof- phenotype. The increase in frameshifting is greatest when the lacZ reporter gene is expressed on a high copy, episomal vector. No differences were found in 5 S rRNA copy number or electrophoretic mobilities in mof9 strains. Both mof9 and rDNA::LEU2 increase the efficiency of +1 frameshifting as well but have no effect on readthrough of UAG or UAA termination codons, indicating that not all translational specificity is affected. These data suggest a role for 5 S rRNA in the maintenance of frame in translation.
可以使用lacZ表达依赖于阅读框 -1 移位的构建体来分离 -1 核糖体移码效率增加的染色体突变体(维持阅读框 = mof)。我们在酿酒酵母中分离到一个新的mof突变体mof9,并表明它可被单拷贝和多拷贝的5 S rDNA克隆互补。rDNA位点的两个独立插入突变(rDNA::LEU2和rDNA::URA3)也表现出Mof-表型,并且也可被单拷贝和多拷贝的5 S rDNA克隆互补。在野生型宿主中,从质粒表达的突变型5 S rRNA占总5 S rRNA的20 - 50%时也会诱导Mof-表型。当lacZ报告基因在高拷贝附加型载体上表达时,移码增加最为显著。在mof9菌株中,未发现5 S rRNA拷贝数或电泳迁移率有差异。mof9和rDNA::LEU2都同样增加了 +1 移码的效率,但对UAG或UAA终止密码子的通读没有影响,这表明并非所有翻译特异性都受到影响。这些数据表明5 S rRNA在翻译中维持阅读框方面发挥作用。