We are sorry, but NCBI web applications do not support your browser and may not function properly. More information

Results: 4

1.
Figure 2

Figure 2. From: A CBF5 mutation that disrupts nucleolar localization of early tRNA biosynthesis in yeast also suppresses tRNA gene-mediated transcriptional silencing.

The mutation leading to suppression of tgm silencing in the CBF5 gene. The gene corresponding to the art1-1 mutation was cloned, and the entire coding region of the parental wild-type and mutant CBF5 genes was sequenced. The mutant contained a single AUG to AUU change in the translation initiation codon.

Ann Kendall, et al. Proc Natl Acad Sci U S A. 2000 November 21;97(24):13108-13113.
2.
Figure 4

Figure 4. From: A CBF5 mutation that disrupts nucleolar localization of early tRNA biosynthesis in yeast also suppresses tRNA gene-mediated transcriptional silencing.

In situ fluorescence of the mutant strain CBF5-AUU and the parental wild-type strain. DAPI stains of the nucleoplasm are blue in all views. The fluorescein-labeled probe to the introns of pre-tRNAs are green, and the Cy3-labeled probe to U14 nucleolar RNA is red. In the wild-type strain, most of the pre-tRNA signal is colocalized to the nucleolus, along with the U14 snoRNA, although there are some secondary loci of pre-tRNA staining. In the CBF5-AUU strain, the U14 nucleolar signal is slightly fragmented, but the pre-tRNA signal is dispersed to the nucleoplasm.

Ann Kendall, et al. Proc Natl Acad Sci U S A. 2000 November 21;97(24):13108-13113.
3.
Figure 1

Figure 1. From: A CBF5 mutation that disrupts nucleolar localization of early tRNA biosynthesis in yeast also suppresses tRNA gene-mediated transcriptional silencing.

(A) Plasmid used to monitor tgm silencing. In this plasmid, the tRNASUP4 gene is placed in the same (s) or opposite (o) orientation as transcription of the HIS3 coding region. HIS3 expression is controlled by a consensus UASGAL and the GAL1 promoter as described previously (1). (B) A Northern blot analysis of HIS3 mRNA and URA3 mRNA, expressed from the reporter plasmid (pSUP4o/s). Both orientations of the neighboring tRNA gene drastically reduce HIS3 mRNA levels.

Ann Kendall, et al. Proc Natl Acad Sci U S A. 2000 November 21;97(24):13108-13113.
4.
Figure 3

Figure 3. From: A CBF5 mutation that disrupts nucleolar localization of early tRNA biosynthesis in yeast also suppresses tRNA gene-mediated transcriptional silencing.

Loss of silencing and slow growth recreated by an AUG to AUU change in the CBF5 initiation codon. (A) The expression of the tRNASUP4 gene and the HIS3 gene on media containing glucose (represses pol II promoter) or galactose (induces pol II promoter). The tRNASUP4 gene is always expressed when present, tested by the suppression of a chromosomal ade2-101 mutation. In the original mutant (art1-1, which is derived from strain YM2062), expression of HIS3 from the GAL1 promoter requires galactose induction. To show that this galactose-inducible His+ phenotype is due solely to the CBF5 AUG to AUU mutation, either the mutated or wild-type CBF5 gene was introduced into an unrelated strain (W3031A) on a plasmid, and the chromosomal CBF5 was deleted. The AUU mutation conferred galactose-inducible HIS3 expression. (B) Recreation of the AUU translational initiation codon in CBF5 (W3031aΔcbf5/pSUP4oCBF5-AUU) also recreates the slow growth phenotype of the original mutant compared with the same construct with the wild-type AUG in the plasmid-borne CBF5 gene.

Ann Kendall, et al. Proc Natl Acad Sci U S A. 2000 November 21;97(24):13108-13113.

Supplemental Content

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...
Write to the Help Desk