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  • 1
    ISSN: 1432-0983
    Keywords: Schizosaccharomyces pombe ; DNA topoisomerases ; rRNA synthesis ; r-Protein turnover
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The synthesis of rRNA and r-proteins was studied in temperature-sensitive topoisomerase mutants of the fisson yeast Schizosaccharomyces pombe. To reduce the severity of heatshock response seen in the wild type strain, slow temperature shift-up of the cultures was used to inactivate the mutant topoisomerases. It was found that the temperature shift caused a large preferential reduction of rRNA synthesis in the top1top2 double mutant. In contrast, no preferential inhibition of rRNA synthesis was observed in top1 or top2 single mutants, although some reduction in the total RNA synthesis was observed in the top2 mutant. Thus, as observed with Saccharomyces cerevisiae (Brill et al. 1987), relaxation of supercoiled DNA structures by either topoisomerase I or II appears to be essential for efficient transcription of rRNA genes. Analysis of r-protein synthesis indicated that there were small decreases in the differential synthesis rates of r-proteins after temperature shift-up in the top1top2 mutant, but the observed negative effects on r-protein synthesis was much smaller than that on rRNA synthesis, and degradation of the newly synthesized r-proteins was observed. These observations indicate the apparent lack of tight coupling between rRNA and r-protein synthesis in S. pombe under these experimental conditions.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1617-4623
    Keywords: Ribosomal protein operon ; Translational repression ; RNA-protein interaction ; Conservation of mRNA structure ; Gene cloning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In Escherichia coli the genes encoding ribosomal proteins L11 (rplK) and L1 (rplA) are contained in a single operon and their expression is translationally regulated by L1. We have cloned the homologous genes from two other enterobacteria, Serratia marcescens and Proteus vulgaris, and determined nucleotide sequences. The genes are organized in a similar way to that found in E. coli. Conservation of nucleotide and amino acid sequences relative to E. coli in the protein coding regions are 89.2% and 94.7% for S. marcescens, and 80.9% and 88.6% for P. vulgaris. Nucleotide sequences of L11 mRNA leader regions were strongly conserved for the primary as well as the secondary structures in the L1 target site. We have also constructed plasmids carrying E. coli L11 and either P. vulgaris or S. marcescens L1 genes fused to the lac promoter, with or without the E. coli leader containing the L1 target site. Induction of transcription of the operons possessing the E. coli mRNA leader did not lead to overproduction of L11, indicating translational regulation of the chimeric operon as well as the chromosomal operon by the plasmid encoded L1. Repression of the chromosomal L11 operon was directly demonstrated upon induction of the chimeric operons without the leader, which also lack the L11 initiation signal but have a mutation allowing L1 translation. These results show that both S. marcescens and P. vulgaris L1, despite differences in the amino acid sequences, can function as a repressor as does E. coli L1, and suggest that, as is the case for the interaction between L1 and rRNA in ribosome assembly, the sites in L1 and mRNA involved in the repressor-mRNA interaction are conserved among these three bacterial species.
    Type of Medium: Electronic Resource
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