ABSTRACT
The E2F family of transcription factors regulates the expression of a number of genes whose products are involved in cell cycle control, DNA replication and apoptosis. We show here that E2F-1 binds in vivo the promoters of ASPP1 and ASPP2 genes, two activators of p53-mediated apoptosis, E2F-1, E2F-2 and E2F-3 all activate the isolated ASPP1 and ASPP2 promoters. Overexpression or deregulation of E2F-1 increased the expression levels of ASPP1 and ASPP2 mRNA and proteins. The identification of ASPP1 and ASPP2 genes as transcriptional targets of E2F provides another mechanism by which E2F cooperates with p53 to induce apoptosis.
Subject(s)
Carrier Proteins/genetics , Cell Cycle Proteins/metabolism , DNA-Binding Proteins/metabolism , Transcription Factors/metabolism , Adaptor Proteins, Signal Transducing , Apoptosis Regulatory Proteins , Base Sequence , Carrier Proteins/biosynthesis , E2F Transcription Factors , E2F1 Transcription Factor , E2F2 Transcription Factor , E2F3 Transcription Factor , Humans , Molecular Sequence Data , Promoter Regions, Genetic , RNA, Messenger/metabolismABSTRACT
Three regulatory genes, known to be required for glucose repression/derepression of some genes in Saccharomyces cerevisiae, were disrupted to study their effects on the carbon-source regulation of the STA2 glucoamylase gene expression. Using a STA2-lacZ fusion it was found that: (1) the MIG1 gene is dispensable for the repression of the STA2 gene; (2) there are two components in the carbon-source repression of STA2: HXK2-dependent and HXK2-independent; and (3) the HAP2 gene seems to be involved in repression rather than activation of the STA2 expression.
Subject(s)
CCAAT-Binding Factor , Gene Expression Regulation, Enzymologic , Gene Expression Regulation, Fungal , Glucan 1,4-alpha-Glucosidase/genetics , Repressor Proteins/genetics , Repressor Proteins/physiology , Saccharomyces cerevisiae/genetics , Alleles , DNA-Binding Proteins/genetics , Fungal Proteins/genetics , Glucose/metabolism , Lac Operon , Saccharomyces cerevisiae Proteins , Transcription Factors/geneticsABSTRACT
A procedure was developed for construction of industrial strains of distiller's yeast (Saccharomyces cerevisiae). It includes several steps: construction of congenic genetically marked haploid strains of opposite mating types starting from an industrial strain of hybrid nature, integrative transformation of the above haploid strains with a DNA fragment containing an expression cassette responsible for new technological facilities, and hybridization of transformants and isolation of final industrial homozygous strains under experimental conditions simulating commercial fermentation processes. This strategy permits the generation of strains that have desirable characteristics of traditional races of distiller's yeast along with new technological facilities determined by the particular expression cassette. Using this procedure, we have constructed an industrial strain with improved amylolytic activity. (c) 1995 John Wiley & Sons, Inc.
ABSTRACT
A diastatic strain of Saccharomyces cerevisiae producing the STA2-encoded extracellular glucoamylase (GA) in a pronounced glucose-repressible fashion was used as a parent for generating mutants with reduced GA activity under normal conditions of derepression. In addition to mutations in STA2, five other recessive mutations were identified which fell into four complementation groups designated haf1 through haf4. RNA blot analysis suggested that the haf mutations confer defects in STA2 transcription. The haf mutants were pleiotropically defective in utilization of alternative carbon sources and resembled the snf (sucrose non-fermenting) mutants identified previously as unable to derepress the expression of the SUC2 gene encoding invertase. We present evidence strongly suggesting that haf1 = snf2, haf3 = snf1 and haf4 = snf5. By phenotypic criteria, the postulated HAF2 gene (which is none of the SNF genes tested) appears to be similar to SNF2, SNF5 and SNF6, and is possibly a non-redundant extension of this group of functionally related SNF genes.
