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This review provides an overview of the current knowledge of ribosome biogenesis, nucleolus structure and function and protein traffic into and out of the nucleus, with emphasis on the potential of yeast Saccharomyces cerevisiae as a model organism.
The newly discovered Saccharomyces cerevisiae gene KRR1 (YCL059c) encodes a protein essential for cell viability. Krr1p contains a motif of clustered basic amino acids highly conserved in the evolutionarly distant species from yeast to human. We demonstrate that Krr1p is localized in the nucleolus. The KRR1 gene is highly expressed in dividing cells and its expression ceases almost completely when cells enter the stationary phase. In vivo depletion of Krr1p leads to drastic reduction of 40S ribosomal subunits due to defective 18S rRNA synthesis. We propose that Krr1p is required for proper processing of pre-rRNA and the assembly of preribosomal 40S subunits.
The Krr1 protein of Saccharomyces cerevisiae is involved in processing of pre-rRNA and assembly of pre-ribosomal 40S subunits. To further investigate the function of Krr1p we constructed a conditional cold sensitive mutant krrl-21, and isolated seven genes from Schizosaccharomyces pombe whose products suppressed the cold sensitive phenotype of krrl-21 cells. Among the multicopy suppressors we found genes coding for translation elongation factor EF-1a, a putative ribose methyltransferase and five genes encoding ribosomal proteins. Using the tandem af­finity purification (TAP) method we identified thirteen S. cerevisiae ribosomal pro­teins interacting with Krr1p. Taken together, these results indicate that Krr1p inter­acts functionally as well as physically with ribosomal proteins. Northern blot analy­sis revealed that changes in the level of krrl-21 mRNA were accompanied by similar changes in the level of mRNAs of genes encoding ribosomal proteins. Thus, Krr1p and the genes encoding ribosomal proteins it interacts with seem to be coordinately regulated at the level of transcription.
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The results of the internal transcribed spacer (ITS2) of extrachromosomal rDNA and the chloroplast SSU rDNA sequence analysis presented here confirmed elevated genetic polymorphism revealed earlier by RFLP and RAPD for seven clones of the cosmopolitan species - Euglena agilis Carter. High diversity among these clonal strains was not reflected by morphological criteria, with the exception of the only one character - the ability of the cell in its non-motile dividing states (palmella) to produce mucus and form a slimy envelope. Evolutionary adaptation as formation of slimy envelope may be attributed to different survival strategy of the species by which it adapts to life in a highly variable environment.
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