Cell
ArticleA quantitative analysis of the effects of 5′ junction and TACTAAC box mutants and mutant combinations on yeast mRNA splicing
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YRA1 Autoregulation Requires Nuclear Export and Cytoplasmic Edc3p-Mediated Degradation of Its Pre-mRNA
2007, Molecular CellCitation Excerpt :We used yra1-N84, a complete loss-of-regulation allele that contains a 252 nt deletion in the coding region of exon 1 and encodes a pre-mRNA that is efficiently spliced, as evidenced by high levels of mRNA but almost no pre-mRNA in wild-type, upf1Δ, edc3Δ, and upf1Δedc3Δ strains (Figure 5C). We found that mutations in the 5′ splice site (m5SS) or the branch point region (mBB2), which block splicing prior to the first step of splicing (Jacquier et al., 1985; Parker and Guthrie, 1985), greatly reduced or eliminated splicing of the yra1-N84 pre-mRNA in wild-type, upf1Δ, edc3Δ, and upf1Δedc3Δ strains (Figure 5C). Remarkably, the m5SS and mBB2 mutations also restored yra1-N84 pre-mRNA to wild-type pre-mRNA regulation; i.e., the yra1-N84-m5SS and yra1-N84-mBB2 pre-mRNAs accumulated to high levels in edc3Δ strains (Figure 5C).
Cotranscriptional spliceosome assembly dynamics and the role of U1 snRNA:5′ss base pairing in yeast
2005, Molecular CellCitation Excerpt :The 5′II and 3′I mutations result in pre-mRNA accumulation as well as 40%–75% and 80%–100% mRNA levels, respectively (Figure 2D; Jacquier et al., 1985). The 3′I mutation is an A to C transversion at position 2 of the BP and results in a ∼5 fold increase in pre-mRNA accumulation, whereas the 5′II construct is a G to A transition at position 5 of the 5′ss and results in a larger ∼15 fold increase in pre-mRNA accumulation (Figure 2D; Jacquier et al., 1985). The U1 pattern is slightly delayed by the 3′I mutation (Figure 5A), and the U2 and U5 patterns show only a modest decrease in peak levels (Figure 5B).
Transformation systems for filamentous fungi and an overview of fungal gene structure
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