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. 2009 May 12;37(13):4205–4217. doi: 10.1093/nar/gkp341

Figure 2.

Figure 2.

Cwc2 is required for cell growth, pre-mRNA splicing, the levels of the U1, U4, U5 and U6 snRNAs and maintenance of the U4/U6 complex. (A) The CWC2 gene was put under control of the GAL1 promoter. Growth over time of the GAL1-CWC2 yeast in the presence of galactose, which allows expression of Cwc2, or glucose, which represses Cwc2 expression. (B) Primer extension analysis of splicing of the U3 intron-containing genes U3A and U3B. When CWC2 is switched off by growth in glucose there is an accumulation of U3 precursor RNA indicating a defect in pre-mRNA splicing. (C) Depletion of Cwc2 influences the levels of the U1, U4, U5 and U6 snRNAs. Primer extension analysis of snRNA levels before and following the switch of the GAL1-CWC2 strain to glucose. The RNA utilized for snRNA primer extension is identical to that used in (B). (D) Depletion of Cwc2 reduces the levels of U4/U6 complex and free U4 and U6. RNA was isolated under nondenaturing conditions from the GAL1-CWC2 strain grown in galactose or glucose, hybridized to radiolabeled probes complementary to U4 or U6 and separated on a 9% polyacrylamide gel.