RESUMEN
The accurate and efficient translation of proteins is of fundamental importance to both bacteria and higher organisms. Most of our knowledge about the control of translational fidelity comes from studies of Escherichia coli. In particular, ram (ribosomal ambiguity) mutations in structural genes of E. coli ribosomal proteins S4 and S5 have been shown to increase translational error frequencies. We describe the first sequence of a ribosomal protein gene that affects translational ambiguity in a eucaryote. We show that the yeast omnipotent suppressor SUP44 encodes the yeast ribosomal protein S4. The gene exists as a single copy without an intron. The SUP44 protein is 26% identical (54% similar) to the well-characterized E. coli S5 ram protein. SUP44 is also 59% identical (78% similar) to mouse protein LLrep3, whose function was previously unknown (D.L. Heller, K.M. Gianda, and L. Leinwand, Mol. Cell. Biol. 8:2797-2803, 1988). The SUP44 suppressor mutation occurs near a region of the protein that corresponds to the known positions of alterations in E. coli S5 ram mutations. This is the first ribosomal protein whose function and sequence have been shown to be conserved between procaryotes and eucaryotes.
Asunto(s)
Escherichia coli/genética , Genes Bacterianos , Genes Fúngicos , Genes Supresores , Proteínas Ribosómicas/genética , Saccharomyces cerevisiae/genética , Secuencia de Aminoácidos , Secuencia de Bases , Clonación Molecular , Prueba de Complementación Genética , Datos de Secuencia Molecular , Mutación , Mapeo Restrictivo , Homología de Secuencia de Ácido NucleicoRESUMEN
Omnipotent suppressors decrease translational fidelity and cause misreading of nonsense codons. In the presence of the non-Mendelian factor [eta+], some alleles of previously isolated omnipotent suppressors are lethal. Thus the current search was conducted in an [eta+] strain in an effort to identify new suppressor loci. A new omnipotent suppressor, SUP39, and alleles of sup35, sup45, SUP44 and SUP46 were identified. Efficiencies of the dominant suppressors were dramatically reduced in strains that were cured of non-Mendelian factors by growth on guanidine hydrochloride. Wild-type alleles of SUP44 and SUP46 were cloned and these clones were used to facilitate the genetic analyses. SUP44 was shown to be on chromosome VII linked to cyh2, and SUP46 was clearly identified as distinct from the linked sup45.
Asunto(s)
Mapeo Cromosómico , Supresión Genética , Levaduras/genética , Alelos , Cromosomas Fúngicos , Clonación Molecular , Cruzamientos Genéticos , Genes Fúngicos , Prueba de Complementación Genética , Ligamiento Genético , Genotipo , Fenotipo , Mapeo RestrictivoRESUMEN
Omnipotent suppressors cause translational ambiguity and have been associated with poor growth and inviability. We now report that a non-Mendelian element, [eta(+)], causes this inviability. In [eta(-)] strains the suppressors are not inviable. The [eta(+)] genetic element segregates to about 70% of the meiotic progeny, although almost all of the spores probably have the [eta(+)] phenotype for the first few divisions. Growth on 5 mM guanidine hydrochloride efficiently causes [eta(+)] strains to become [eta(-)]. The [eta(+)] factor has many similarities with the previously described [psi(+)] factor (Cox 1965, 1971). However, [eta(+)] and [psi(+)] differ in their patterns of inheritance, and by the fact that [psi(+)] affects ochre specific and not omnipotent suppressors, while the converse is true of [eta(+)].