The upstream region of the isocitrate lyase gene (UPR-ICL) of Candida tropicalis induces gene expression in both Saccharomyces cerevisiae and Escherichia coli by acetate via two distinct promoters

Atomi, Haruyuki; Umemura, Ken; Higashijima, Takanori; Kanai, Tamotsu; Yotsumoto, Yoshihisa; Teranishi, Yutaka; Ueda, Mitsuyoshi; Tanaka, Atsuhiro

doi:10.1007/s002030050210

Cited by 3 publications

(1 citation statement)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Second, the UPR – ICL promoter was used instead of the intrinsic promoters to construct expression plasmids. The UPR – ICL promoter was derived from Candida tropicalis and was strong and functional under the decrease of glucose concentration in the culture medium (Atomi et al ., 1995; Umemura et al ., 1995a, b). The expression of the constructed Aga1p–Aga2p fusion protein was responsive to the change of the concentration of glucose in the medium without exposure to the opposite mating type cells and the protein expressed was proved to be functional toward Agα1p.…”

Section: Introductionmentioning

confidence: 99%

Establishment of a simple system to analyse the molecular interaction in the agglutination ofSaccharomyces cerevisiae

Zou

Ueda

Murai

et al. 2000

Yeast

Self Cite

View full text Add to dashboard Cite

Saccharomyces cerevisiae a‐agglutinin, which is involved in mating and covalently anchoring to the cell wall, consists of two components, Aga1p and Aga2p, whose syntheses are individually regulated. To facilitate the analysis of the protein–protein interaction on agglutination between a‐ and α‐agglutinins, the construction of a yeast strain (MATa) with the functional protein prepared by genetic fusion of Aga1p‐ and Aga2p‐encoding genes and by the expression system using the UPR–ICL promoter derived from the n‐alkane‐assimilating yeast, Candida tropicalis, which is functional under the condition of lower glucose concentration was tried and the agglutination ability of the constructed strain was evaluated with a yeast strain (MATa) which expressed AGα1 encoding α‐agglutinin under the control of the same promoter. The genes were integrated into the yeast chromosomes. Cell agglutination between both (MATa) strains was observed microscopically when these two strains were mix‐cultured to a glucose‐decreased concentration. The agglutination was further confirmed by the sedimentation test and by the quantification using a filter. These results proved that the constructed Aga1p–Aga2p fusion protein was enoughly functional for the interaction with the Agα1 protein, and that this phenomenon occurred dependent on glucose concentration, but independent of the peptide pheromones secreted by the cells of the opposite mating types. Using this system, the role of two disulphide linkages between Aga1p and Aga2p on the binding activity between Aga2p and Aga1p was first evaluated. Under the treatment by the SH‐compound (dithiothreitol), in which Agα2p is easily released into the medium from the intact cell surface, the Aga1p and Aga2p fusion protein was a good tool to make clear the role of the disulphide linkages. As a result, the linkages had a significant effect on not only the assembly but also the binding activity. The novel and simple system described here may further facilitate the study of molecular interaction in agglutination. Copyright © 2000 John Wiley & Sons, Ltd.

show abstract

Section: Introductionmentioning

confidence: 99%

Establishment of a simple system to analyse the molecular interaction in the agglutination ofSaccharomyces cerevisiae

Zou

Ueda

Murai

et al. 2000

Yeast

Self Cite

View full text Add to dashboard Cite

show abstract

Derepression of Gene Expression Mediated by the 5′ Upstream Region of the Isocitrate Lyase Gene of Candida Tropicalis is Controlled by two Distinct Regulatory Pathways in Saccharomyces Cerevisiae

Umemura

Atomi

Kanai

et al. 1997

European Journal of Biochemistry

Self Cite

View full text Add to dashboard Cite

The 5' upstream region of the gene encoding isocitrate lyase of Cundida tropicalis (UPR-ICL) is functional as a promoter in Saccharomyces cerevisiae, and it is regulated by carbon source; the expression of the gene is repressed when cells are grown on glucose, while it increases to a higher level in acetategrown cells. Therefore, we have investigated regions in UPR-ICL responsible for gene expression in glucose-grown and acetate-grown cells. In glucose-grown cells, a deletion of the region between -801 and -569 (region G1) significantly decreased gene expression compared with that observed with the complete UPR-ICL. The region from -421 to -379 (region G2) also repressed gene expression in glucose-grown cells. In acetate-grown cells, two regions were found to strongly enhance gene expression, one between -728 and -569 (region A l ) and the other between -370 and -356 (region A2). Whereas region A2 contained a sequence motif similar to the carbon-source-responsive element (CSRE), which mediates regulation by carbon source of S. cerevisiae ICLl, region A1 did not show similarity to any reported cis-acting elements. Deletion mutants of UPR-ICL containing only one of these regions showed that each region could independently activate gene expression to a similar level when the cells were grown on acetate. The influences of null mutations in the MIGI, SNFI and CAT8 genes on regulation of UPR-ICL-mediated gene expression were examined. Expression of the ICL gene with full-length UPR-ICL increased about tenfold in migl cells grown on glucose, while little difference was observed in acetate-grown cells. The effects of snfI and cut8 mutations were different between region-Al-mediated and region-A2-mediated gene expression in acetate-grown cells. Region-AZmediated expression decreased 9. 5 % and 84% in snfI and cat8 cells, respectively, while region-Al-mediated expression decreased 72% in snfI cells and was not affected by the cat8 mutation. This finding indicates that regionAl-mediated gene expression is regulated by a pathway independent of CAT8, which is necessary for derepression of CSRE-mediated gene expression in S. cerevisiue Keywords: upstream region of the isocitrate lyase gene; glucose repression ; isocitrate lyase ; Succharomyces cerevisiae; Cundida tropicalis.Glucose repression is the process by which the synthesis of enzymes participating in utilization of carbon sources other than glucose is repressed in the presence of glucose. In the yeast Saccharomyces cerevisiae, the genes encoding the enzymes for alternative-sugar utilization, the glyoxylate cycle and gluconeogenesis are under this regulation, and its mechanism has been investigated extensively [ 1 -51. Investigations of cis-acting elements have been conducted in the upstream regions of the genes encoding fructose 1,6-bisphosphatase ( F B P I ) [6-81 and phosphoenolpyruvate carboxykinase (PCKl) [9 j of the gluconeogenic pathway, and isocitrate lyase (ICL1) [lo] of the glyoxylate cycle. In ICLl, a positive element called the carbonsource-responsive eleme...

show abstract

Transcriptional Regulation of Peroxisomal Glyoxylate Cycle Enzymes of an n‐Alkane‐Assimilating Yeast, Candida tropicalis

Atomi

Umemura

Ueda

et al. 1996

Annals of the New York Academy of Sciences

View full text Add to dashboard Cite

The upstream region of the isocitrate lyase gene (UPR-ICL) of Candida tropicalis induces gene expression in both Saccharomyces cerevisiae and Escherichia coli by acetate via two distinct promoters

Cited by 3 publications

References 0 publications

Establishment of a simple system to analyse the molecular interaction in the agglutination ofSaccharomyces cerevisiae

Establishment of a simple system to analyse the molecular interaction in the agglutination ofSaccharomyces cerevisiae

Derepression of Gene Expression Mediated by the 5′ Upstream Region of the Isocitrate Lyase Gene of Candida Tropicalis is Controlled by two Distinct Regulatory Pathways in Saccharomyces Cerevisiae

Transcriptional Regulation of Peroxisomal Glyoxylate Cycle Enzymes of an n‐Alkane‐Assimilating Yeast, Candida tropicalis

Contact Info

Product

Resources

About