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. 2009 Mar 26;28(9):1260–1270. doi: 10.1038/emboj.2009.79

Figure 6.

Figure 6

Identification of a novel snR30 element required for 18S processing. (A) Structure and expression of snR30 RNAs carrying altered 3′-terminal hairpins. Sequences used to replace wild-type nucleotides in the pR30 expression construct are highlighted in shaded boxes. The resulting pR30st1, pR30st2 and pR30-R5 expression plasmids were transformed into yeast GAL∷snR30 cells and expression of the mutant snR30 RNAs was monitored by northern blot analysis. (B) Northern blot analysis of pre-rRNA processing in GAL∷snR30 cells transformed with the indicated expression plasmids. (C) Structure and expression of mutant snR30-R5d, snR30-R5p, snR30hp and snR30lm RNAs in GAL∷snR30 cells. (D) Processing of pre-rRNA in GAL∷snR30 cells transformed with the indicated plasmids. (E) Sedimentation of R30-R5d snoRNA in sucrose gradient. Extracts of GAL∷snR30 cells expressing R30-R5d or R30 snoRNAs were fractionated in a 4.5–45% sucrose gradient. RNA was isolated from each of 18 fractions and analysed by northern blotting with R30- and 35S-specific oligonucleotide probes. Positions of 40S, 60S and 80S ribosomal particles are indicated.