Journal ArticleDOI
Mutational analysis of the upstream activation site of yeast ribosomal protein genes.
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TLDR
To investigate the sequence requirements of the RPG box, synthetic oligonucleotides were inserted into a deletion mutant of the L25 promoter that lacked its natural RPG elements and showed that in the 3′ part of this sequence element single substitutions are allowed at all positions, in the 5′ part, however, the nucleotide requirements appear to be more stringent.Abstract:
Most ribosomal protein (rp-)genes in yeast are preceded by conserved sequence motifs that act as upstream transcription-activating sites (RPG box). These sequence elements have previously been shown to represent specific binding sites for a protein factor, TUF. Comparison of the various nucleotide elements identified so far indicates a remarkably high degree of variation in the respective sequences. On the other hand, a methylation interference study performed with one RPG box revealed close contact points with the TUF protein along the entire sequence. To investigate the sequence requirements of the RPG box, we inserted synthetic oligonucleotides that differed from the general consensus sequence ACACCCATACATTT at single positions into a deletion mutant of the L25 promoter that lacked its natural RPG elements. Transcription activity was estimated by Northern analyses of the cellular level of L25-galK hybrid transcripts. The results show that in the 3′ part of this sequence element single substitutions are allowed at all positions, in the 5′ part, however, the nucleotide requirements appear to be more stringent. In particular, the invariant C at position 5 of the consensus sequence is absolutely necessary for its enhancer function.read more
Citations
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Journal ArticleDOI
RAP1: a protean regulator in yeast
TL;DR: The yeast protein RAP1 is a sequence-specific DNA-binding protein that binds to many promoters, to two elements that silence mating-type genes, and to [(C)1-3A]n tracts at telomeres.
Journal ArticleDOI
Introduction and expression of genes for metabolic engineering applications in Saccharomyces cerevisiae
TL;DR: This MiniReview focuses on methods for introducing genes and controlling their copy number and on the many promoters (both constitutive and inducible) that have been successfully employed.
Journal ArticleDOI
Central role of Ifh1p–Fhl1p interaction in the synthesis of yeast ribosomal proteins
TL;DR: Evidence is provided that two factors, Fhl1p and Ifh1p, are key players in the transcription of RP genes, both found at transcribing RP genes in vivo and both severely deleterious to ribosome synthesis and cell growth.
Journal ArticleDOI
Association of RAP1 binding sites with stringent control of ribosomal protein gene transcription in Saccharomyces cerevisiae.
C M Moehle,A G Hinnebusch +1 more
TL;DR: In this paper, the authors used the antimetabolite 3-amino-1,2,4-triazole to cause stringent control in the yeast Saccharomyces cerevisiae.
Journal ArticleDOI
Transcriptional elements involved in the repression of ribosomal protein synthesis.
TL;DR: It is concluded that the cell has evolved a two-pronged approach to effect the rapid extinction of RP synthesis in response to the stress imposed by a heat shock or by a failure of the secretory pathway.
References
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Journal ArticleDOI
DNA sequencing with chain-terminating inhibitors
TL;DR: A new method for determining nucleotide sequences in DNA is described, which makes use of the 2',3'-dideoxy and arabinon nucleoside analogues of the normal deoxynucleoside triphosphates, which act as specific chain-terminating inhibitors of DNA polymerase.
Book ChapterDOI
Sequencing end-labeled DNA with base-specific chemical cleavages.
Allan M. Maxam,Walter Gilbert +1 more
TL;DR: The chapter presents techniques for producing discrete DNA fragments, end-labeling DNA, segregating end- labeled fragments, extracting DNA from gels, and the protocols for partially cleaving it at specific bases using the chemical reactions.
Journal ArticleDOI
Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange
TL;DR: A simple and rapid system for the denaturation of nucleic acids and their subsequent analysis by gel electrophoresis, using the metachromatic stain acridine orange for visualization ofucleic acids in gels and which can be analyzed on the same slab gel.
Journal ArticleDOI
Purification and cloning of a DNA binding protein from yeast that binds to both silencer and activator elements
David Shore,Kim Nasmyth +1 more
TL;DR: The purification of RAP1 by DNA affinity chromatography and the isolation of its gene from a lambda gt11 genomic library using antibodies raised against the protein suggest that R AP1 may be a transcriptional regulator that can play a role in either repression or activation of transcription, depending upon the context of its binding site.
Journal ArticleDOI
The making of strand-specific M13 probes.
Hu Nien-tai,Joachim Messing +1 more
TL;DR: A novel approach has been developed for the preparation of highly radioactive, strand-specific M13 probes by using a universal primer to initiate the DNA synthesis of the complementary strand of the M13 sequence downstream from the inserted sequence.
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