Analysis of Expression of Yeast Enolase 1 Gene Containing a Longer Pyrimidine-Rich Region Located between the TATA Box and Transcription Start Site

Abstract

Yeast ENO1 promoter was prepared by a chemical synthetic method. Two variant promoters containing a pyrimidine-rich region (CT block), located between the TATA box and the transcription start site, either 32 or 34 base pairs (bp) longer than the native ENO1 promoter were isolated during the chemical synthesis. Gene expression of variant promoters was compared with that of the native promoter by measuring the amount of mRNA and the activity of beta-galactosidase by constructing ENO1-lacZ gene fusions. No significant differences were observed between the native and variant promoters in transcription levels. The start site of transcription was mapped on CAAG, a consensus sequence of transcription start site of yeast glycolytic genes. The results suggest a longer CT block in ENO1 promoter may not affect the expression of the yeast ENO1 gene. In addition, the level of ENO1 gene expression was found to be higher in stationary phase cells than in log phase cells.

Enolase from the ectomycorrhizal fungus Tuber borchii Vittad.: biochemical characterization, molecular cloning, and localization

“…The purification fold and yield value obtained were higher than most of those reported for other organisms (Jigami et al, 1986;Peak et al, 1994).…”

“…The presence of only one gene has been highlighted in other organisms such as C. elegans and A. oryzae (Machida et al, 1996;Wang et al, 2000) although in most fungi the presence of multiple enolase genes have been described. In S. cerevisiae two enolase genes, ENO1 and ENO2 (Holland et al, 1981), are present and they are differentially regulated by the cell growth phase (Jigami et al, 1986;McAlister and Holland, 1982) and by the carbon source used to propagate the cells (Cohen et al, 1986;Maitra and Lobo, 1971). The same regulation has been demonstrated also in A. oryzae, where only one enolase gene is present (Nakajima et al, 2000).…”

Functional characterization of a pyrimidine-rich element in the 5'-noncoding region of the yeast iso-1-cytochrome c gene.

“…These results may suggest that transcriptional inhibition is due to alteration in the wild-type spatial relationship between functional CYC) TATA elements and corresponding transcription start sites, rather than direct effects due to the presence or absence of the pyrimidine-rich element. In other studies an increase in the length of the yeast ENO] pyrimidine-rich sequence, located between a TATA element and corresponding transcription start sites, by 32 and 34 bp did not affect the level or start sites of transcription (10). Thus, the VOL.…”

The Expression of the γ-Glutamylcysteine Synthetase Gene ofEscherichia coliB inSaccharomyces cerevisiae

“…were mapped at the sequence CAAG. 33 ) Since the P8 sequence had a CT block of about 70 bp, between the TAT A box and the mRNA start site and the mRNA start sites of P8 were mapped at and around a CAAA sequence, it was anticipated that P8 was an efficient transcriptional promoter.…”