Efficient expression of the Saccharomyces cerevisiae glycolytic gene ADH1 is dependent upon a cis -acting regulatory element (UASRPG) found initially in genes encoding ribosomal proteins

Tornow, Joanne; Santangelo, George M.

doi:10.1016/0378-1119(90)90441-s

Cited by 64 publications

(44 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Rap1p sites are found in promoters of ribosomal and glycolytic genes in either orientation and, especially within ribosomal promoters, in multiple copies (Woudt et al 1987;Tornow and Santangelo 1990). Surprisingly, the inverted Rap1p site had no effect on GFP expression ( Figure 5A).…”

Section: Down-regulation Depends On Orientation and Number Of Sitesmentioning

confidence: 97%

“…The resulting synthetic codon-optimized Ty1 (CO-Ty1) was defective for Ty1 transposition and had reduced Ty1 protein and fulllength RNA levels even though the known Ty1 cis-sequences had not been altered, suggesting identification of a novel cisacting sequence capable of regulating Ty1 expression. Mapping and testing of this cis-acting sequence, however, revealed that the defect was the consequence of an inhibitory sequence in CO-Ty1 in the form of a Rap1p binding site.Rap1p (Repressor Activator Protein) is a multifunctional protein with the binding site consensus ACACCCRYACAYM (Lieb et al 2001) with transcriptionally opposing roles in the activation of transcription at promoters of ribosomal and glycolytic genes (Woudt et al 1987;Tornow and Santangelo 1990) as well as in transcriptional silencing through the recruitment of silencing factors to the HM loci and telomeres (Shore and Nasmyth 1987;Buchman et al 1988;Kyrion et al 1992). The latter activity has been attributed to the nonessential Rap1-S domain, which is separate from the DNA binding domain (Sussel and Shore 1991;Kyrion et al 1992).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Novel Transcript Truncating Function of Rap1p Revealed by Synthetic Codon-Optimized Ty1 Retrotransposon

et al. 2012

View full text Add to dashboard Cite

Extensive mutagenesis via massive recoding of retrotransposon Ty1 produced a synthetic codon-optimized retrotransposon (CO-Ty1). CO-Ty1 is defective for retrotransposition, suggesting a sequence capable of down-regulating retrotransposition. We mapped this sequence to a critical 20-bp region within CO-Ty1 reverse transcriptase (RT) and confirmed that it reduced Ty1 transposition, protein, and RNA levels. Repression was not Ty1 specific; when introduced immediately downstream of the green fluorescent protein (GFP) stop codon, GFP expression was similarly reduced. Rap1p mediated this down-regulation, as shown by mutagenesis and chromatin immunoprecipitation. A regular threefold drop is observed in different contexts, suggesting utility for synthetic circuits. A large reduction of RNAP II occupancy on the CO-Ty1 construct was observed 39 to the identified Rap1p site and a novel 39 truncated RNA species was observed. We propose a novel mechanism of transcriptional regulation by Rap1p whereby it serves as a transcriptional roadblock when bound to transcription unit sequences. SACCHAROMYCES cerevisiae Ty1 is the best-characterized and most abundant of five retrotransposon families present in the genome. Ty1 shares many similarities with retroviruses, such as HIV-1 and makes similar use of an RNA intermediate in its propagation. Unsurprisingly, this Ty1 RNA intermediate is essential for Ty1 retrotransposition. Not only does this "mRNA" code for the GAG and POL proteins required for retrotransposition, but it also serves a critical function as the genetic material for replication. Both Ty1 mRNA and its cDNA product contain cis-acting nucleotide determinants required for retrotransposition.Mutagenesis of Ty1 and screening for cis-acting nucleotide determinants has been previously performed with various strategies, including in-frame linker insertion mutagenesis Devine and Boeke 1994;Monokian et al. 1994), analysis of synthetic Ty1 DNA fragments to determine minimal requirements for the integration reaction (Eichinger and Boeke 1990;Braiterman et al. 1994;Devine and Boeke 1994;Sharon et al. 1994), deletion analysis and PCR-mediated mutagenesis of mini-Ty1 elements (Xu and Boeke 1990;Bolton et al. 2005), and comparative sequence analysis followed by targeted mutagenesis and/or generation of complementary mutations in RNA stem regions (Chapman et al. 1992;Heyman et al. 1995;Lauermann et al. 1995;Friant et al. 1998;Cristofari et al. 2002;Bolton et al. 2005). These studies provided insights into mechanisms underlying Ty1 retrotransposition and revealed several cis-acting sequences, including the Met i -tRNA:primer binding site (PBS) interaction (Chapman et al. 1992; Friant et al. 1998), LTRs (Eichinger andBraiterman et al. 1994;Devine and Boeke 1994;Sharon et al. 1994), polypurine tracts (PPTs) (Heyman et al. 1995;Lauermann et al. 1995), the GAG-POL frameshift region (Belcourt and Farabaugh 1990; Lawler et al. 2001), CYC5 and CYC3 (Cristofari et al. 2002), and an extensive 59 RNA structure required for efficient ini...

show abstract

Section: Down-regulation Depends On Orientation and Number Of Sitesmentioning

confidence: 97%

mentioning

confidence: 99%

Novel Transcript Truncating Function of Rap1p Revealed by Synthetic Codon-Optimized Ty1 Retrotransposon

et al. 2012

View full text Add to dashboard Cite

show abstract

“…A wealth of additional evidence suggests that the Rap1/Gcr1 assemblage participates directly in an activation mechanism shared by this common set of target genes. First, Gcr1 activation of glycolytic and RP genes is eliminated by point mutations or small deletions that eliminate the Rap1 binding site(s) in the corresponding promoters (374,375). Thus, the capacity of Gcr1 to stimulate transcription of most if not all of its target genes is entirely dependent upon Rap1 binding upstream of those genes.…”

Section: Transcriptional Induction Of Glycolytic and Ribosomal Proteimentioning

confidence: 99%

Glucose Signaling in Saccharomyces cerevisiae

Santangelo

2006

Microbiol Mol Biol Rev

Self Cite

479

481

View full text Add to dashboard Cite

SUMMARY Eukaryotic cells possess an exquisitely interwoven and fine-tuned series of signal transduction mechanisms with which to sense and respond to the ubiquitous fermentable carbon source glucose. The budding yeast Saccharomyces cerevisiae has proven to be a fertile model system with which to identify glucose signaling factors, determine the relevant functional and physical interrelationships, and characterize the corresponding metabolic, transcriptomic, and proteomic readouts. The early events in glucose signaling appear to require both extracellular sensing by transmembrane proteins and intracellular sensing by G proteins. Intermediate steps involve cAMP-dependent stimulation of protein kinase A (PKA) as well as one or more redundant PKA-independent pathways. The final steps are mediated by a relatively small collection of transcriptional regulators that collaborate closely to maximize the cellular rates of energy generation and growth. Understanding the nuclear events in this process may necessitate the further elaboration of a new model for eukaryotic gene regulation, called “reverse recruitment.” An essential feature of this idea is that fine-structure mapping of nuclear architecture will be required to understand the reception of regulatory signals that emanate from the plasma membrane and cytoplasm. Completion of this task should result in a much improved understanding of eukaryotic growth, differentiation, and carcinogenesis.

show abstract

“…Nevertheless, their inherent regulation modes and fluctuations in expression must be taken into account when choosing a promoter for a given application. For example, the high activity of the original ADH1 promoter depends on the presence of a fermentable carbon source (352). Indeed, there have been multiple attempts to optimize the ADH1 promoter for either brewing fermentation or heterologous protein production (257,299).…”

Section: Changing Protein Cellular Levelsmentioning

confidence: 99%

Progress in Metabolic Engineering of Saccharomyces cerevisiae

Nevoigt

2008

Microbiol Mol Biol Rev

526

333

View full text Add to dashboard Cite

SUMMARY The traditional use of the yeast Saccharomyces cerevisiae in alcoholic fermentation has, over time, resulted in substantial accumulated knowledge concerning genetics, physiology, and biochemistry as well as genetic engineering and fermentation technologies. S. cerevisiae has become a platform organism for developing metabolic engineering strategies, methods, and tools. The current review discusses the relevance of several engineering strategies, such as rational and inverse metabolic engineering, evolutionary engineering, and global transcription machinery engineering, in yeast strain improvement. It also summarizes existing tools for fine-tuning and regulating enzyme activities and thus metabolic pathways. Recent examples of yeast metabolic engineering for food, beverage, and industrial biotechnology (bioethanol and bulk and fine chemicals) follow. S. cerevisiae currently enjoys increasing popularity as a production organism in industrial (“white”) biotechnology due to its inherent tolerance of low pH values and high ethanol and inhibitor concentrations and its ability to grow anaerobically. Attention is paid to utilizing lignocellulosic biomass as a potential substrate.

show abstract

Efficient expression of the Saccharomyces cerevisiae glycolytic gene ADH1 is dependent upon a cis -acting regulatory element (UASRPG) found initially in genes encoding ribosomal proteins

Cited by 64 publications

References 40 publications

Novel Transcript Truncating Function of Rap1p Revealed by Synthetic Codon-Optimized Ty1 Retrotransposon

Novel Transcript Truncating Function of Rap1p Revealed by Synthetic Codon-Optimized Ty1 Retrotransposon

Glucose Signaling in Saccharomyces cerevisiae

Progress in Metabolic Engineering of Saccharomyces cerevisiae

Contact Info

Product

Resources

About