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| Proteins, Expression, Isolation and Analysis | … Then supernatant was collected and incubated with Ni-NTA agarose beads (Genscript) for 1 h at 4C. The beads were washed three times with 10 ml ice-cold washing buffer (20 mM Tris–HCl, pH 7.5, 150 mM KCl, 1 mM MgCl 2 , 1 mM PMSF and 50 mM imidazole) to remove … | Get A Quote |
Ribosomes of different species share an evolutionarily conserved core, exhibiting flexible shells formed partially by the addition of species-specific ribosomal RNAs (rRNAs) with largely unexplored functions. In this study, we showed that by swapping the Saccharomyces cerevisiae 25S rRNA genes with non-S. cerevisiae homologs, species-specific rRNA variations caused moderate to severe pre-rRNA processing defects. Specifically, rRNA substitution by the Candida albicans caused severe growth defects and deficient pre-rRNA processing. We observed that such defects could be attributed primarily to variations in expansion segment 7L (ES7L) and could be restored by an assembly factor Noc2p mutant (Noc2p-K384R). We show... More
