scholarly journals Complex transcriptional regulation of the Saccharomyces cerevisiae CYB2 gene encoding cytochrome b2: CYP1(HAP1) activator binds to the CYB2 upstream activation site UAS1-B2.

1991 ◽  
Vol 11 (7) ◽  
pp. 3762-3772 ◽  
Author(s):  
T Lodi ◽  
B Guiard
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Regulatory Region ◽  
Transcriptional Regulator ◽  
Direct Analysis ◽  
Transcriptional Level ◽  
Glucose Fermentation ◽  
Gene Encoding ◽  
Dnase I Footprinting ◽  
Mrna Transcripts ◽  
Binding Sequence

Expression of the Saccharomyces cerevisiae gene encoding cytochrome b2 (EC 1.2.2.3), CYB2, was investigated by direct analysis of mRNA transcripts and by measurement of the expression of lacZ fused to the CYB2 control regions. These studies indicated that regulation of the CYB2 gene is subject to several metabolic controls at the transcriptional level: inhibition due to glucose fermentation, induction by lactate, and inhibition in anaerobiosis or in absence of heme biosynthesis. Furthermore, we have shown that the CYB2 promoter contains one cis negative regulatory region and two heme-dependent positive regions, one of which is controlled by the transcriptional regulator CYP1 (HAP1) which is involved in the modulation of the expression of several oxygen-regulated genes. The CYP1 (HAP1)-binding sequence was located by gel retardation and DNase I footprinting experiments and compared with the binding sequences previously characterized in detail (UAS1CYC1, UAS'CYP3 (CYC7), and UASCTT1).

Complex transcriptional regulation of the Saccharomyces cerevisiae CYB2 gene encoding cytochrome b2: CYP1(HAP1) activator binds to the CYB2 upstream activation site UAS1-B2

1991 ◽  
Vol 11 (7) ◽  
pp. 3762-3772
Author(s):  
T Lodi ◽  
B Guiard
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Regulatory Region ◽  
Transcriptional Regulator ◽  
Direct Analysis ◽  
Transcriptional Level ◽  
Glucose Fermentation ◽  
Gene Encoding ◽  
Dnase I Footprinting ◽  
Mrna Transcripts ◽  
Binding Sequence

Expression of the Saccharomyces cerevisiae gene encoding cytochrome b2 (EC 1.2.2.3), CYB2, was investigated by direct analysis of mRNA transcripts and by measurement of the expression of lacZ fused to the CYB2 control regions. These studies indicated that regulation of the CYB2 gene is subject to several metabolic controls at the transcriptional level: inhibition due to glucose fermentation, induction by lactate, and inhibition in anaerobiosis or in absence of heme biosynthesis. Furthermore, we have shown that the CYB2 promoter contains one cis negative regulatory region and two heme-dependent positive regions, one of which is controlled by the transcriptional regulator CYP1 (HAP1) which is involved in the modulation of the expression of several oxygen-regulated genes. The CYP1 (HAP1)-binding sequence was located by gel retardation and DNase I footprinting experiments and compared with the binding sequences previously characterized in detail (UAS1CYC1, UAS'CYP3 (CYC7), and UASCTT1).

scholarly journals Regulation ofperRExpression by Iron and PerR in Campylobacter jejuni

2011 ◽  
Vol 193 (22) ◽  
pp. 6171-6178 ◽  
Author(s):  
Minkyeong Kim ◽  
Sunyoung Hwang ◽  
Sangryeol Ryu ◽  
Byeonghwa Jeon
Keyword(s):  
Oxidative Stress ◽  
Campylobacter Jejuni ◽  
Regulatory Region ◽  
Transcriptional Level ◽  
Mobility Shift ◽  
Content Type ◽  
Dnase I Footprinting ◽  
Transcriptional Changes ◽  
Electrophoretic Mobility Shift Assays ◽  
Binding Sequence

Campylobacter jejuniis a leading food-borne pathogen causing gastroenteritis in humans. Although OxyR is a widespread oxidative stress regulator in many Gram-negative bacteria,C. jejunilacks OxyR and instead possesses the metalloregulator PerR. Despite the important role played by PerR in oxidative stress defense, little is known about the factors influencingperRexpression inC. jejuni. In this study, aperRpromoter-lacZfusion assay demonstrated that iron significantly reduced the level ofperRtranscription, whereas other metal ions, such as copper, cobalt, manganese, and zinc, did not affectperRtranscription. Notably, aperRmutation substantially increased the level ofperRtranscription and intranscomplementation restored the transcriptional changes, suggestingperRis transcriptionally autoregulated inC. jejuni. In theperRmutant, iron did not repressperRtranscription, indicating the iron dependence ofperRexpression results fromperRautoregulation. Electrophoretic mobility shift assays showed that PerR binds to theperRpromoter, and DNase I footprinting assays identified a PerR binding site overlapping the −35 region of the twoperRpromoters, further supportingperRautoregulation at the transcriptional level. Alignment of the PerR binding sequence in theperRpromoter with the regulatory region of other PerR regulon genes ofC. jejunirevealed a 16-bp consensus PerR binding sequence, which shares high similarities to theBacillus subtilisPerR box. The results of this study demonstrated that PerR directly interacts with theperRpromoter and regulatesperRtranscription and thatperRautoregulation is responsible for the repression ofperRtranscription by iron inC. jejuni.

scholarly journals Self-regulation of 70-kilodalton heat shock proteins in Saccharomyces cerevisiae.

1990 ◽  
Vol 10 (4) ◽  
pp. 1622-1632 ◽  
Author(s):  
D E Stone ◽  
E A Craig
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Heat Shock ◽  
Heat Shock Proteins ◽  
Self Regulation ◽  
Regulatory Region ◽  
Transcriptional Level ◽  
Transcriptional Fusion ◽  
Heat Shock Element ◽  
Upstream Activating Sequence ◽  
Shock Proteins

To determine whether the 70-kilodalton heat shock proteins of Saccharomyces cerevisiae play a role in regulating their own synthesis, we studied the effect of overexpressing the SSA1 protein on the activity of the SSA1 5'-regulatory region. The constitutive level of Ssa1p was increased by fusing the SSA1 structural gene to the GAL1 promoter. A reporter vector consisting of an SSA1-lacZ translational fusion was used to assess SSA1 promoter activity. In a strain producing approximately 10-fold the normal heat shock level of Ssa1p, induction of beta-galactosidase activity by heat shock was almost entirely blocked. Expression of a transcriptional fusion vector in which the CYC1 upstream activating sequence of a CYC1-lacZ chimera was replaced by a sequence containing a heat shock upstream activating sequence (heat shock element 2) from the 5'-regulatory region of SSA1 was inhibited by excess Ssa1p. The repression of an SSA1 upstream activating sequence by the SSA1 protein indicates that SSA1 self-regulation is at least partially mediated at the transcriptional level. The expression of another transcriptional fusion vector, containing heat shock element 2 and a lesser amount of flanking sequence, is not inhibited when Ssa1p is overexpressed. This suggests the existence of an element, proximal to or overlapping heat shock element 2, that confers sensitivity to the SSA1 protein.

scholarly journals Subcellular Localization of Fad1p in Saccharomyces cerevisiae: A Choice at Post-Transcriptional Level?

Life ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 967 ◽  
Author(s):  
Francesco Bruni ◽  
Teresa Anna Giancaspero ◽  
Mislav Oreb ◽  
Maria Tolomeo ◽  
Piero Leone ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protein Targeting ◽  
Bioinformatic Analysis ◽  
Protein Product ◽  
Transcriptional Level ◽  
Mitochondrial Localization ◽  
Gene Encoding ◽  
Cis Acting ◽  
Per Se ◽  
Shed Light

FAD synthase is the last enzyme in the pathway that converts riboflavin into FAD. In Saccharomyces cerevisiae, the gene encoding for FAD synthase is FAD1, from which a sole protein product (Fad1p) is expected to be generated. In this work, we showed that a natural Fad1p exists in yeast mitochondria and that, in its recombinant form, the protein is able, per se, to both enter mitochondria and to be destined to cytosol. Thus, we propose that FAD1 generates two echoforms—that is, two identical proteins addressed to different subcellular compartments. To shed light on the mechanism underlying the subcellular destination of Fad1p, the 3′ region of FAD1 mRNA was analyzed by 3′RACE experiments, which revealed the existence of (at least) two FAD1 transcripts with different 3′UTRs, the short one being 128 bp and the long one being 759 bp. Bioinformatic analysis on these 3′UTRs allowed us to predict the existence of a cis-acting mitochondrial localization motif, present in both the transcripts and, presumably, involved in protein targeting based on the 3′UTR context. Here, we propose that the long FAD1 transcript might be responsible for the generation of mitochondrial Fad1p echoform.

Self-regulation of 70-kilodalton heat shock proteins in Saccharomyces cerevisiae

1990 ◽  
Vol 10 (4) ◽  
pp. 1622-1632
Author(s):  
D E Stone ◽  
E A Craig
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Heat Shock ◽  
Heat Shock Proteins ◽  
Self Regulation ◽  
Regulatory Region ◽  
Transcriptional Level ◽  
Transcriptional Fusion ◽  
Heat Shock Element ◽  
Upstream Activating Sequence ◽  
Shock Proteins

To determine whether the 70-kilodalton heat shock proteins of Saccharomyces cerevisiae play a role in regulating their own synthesis, we studied the effect of overexpressing the SSA1 protein on the activity of the SSA1 5'-regulatory region. The constitutive level of Ssa1p was increased by fusing the SSA1 structural gene to the GAL1 promoter. A reporter vector consisting of an SSA1-lacZ translational fusion was used to assess SSA1 promoter activity. In a strain producing approximately 10-fold the normal heat shock level of Ssa1p, induction of beta-galactosidase activity by heat shock was almost entirely blocked. Expression of a transcriptional fusion vector in which the CYC1 upstream activating sequence of a CYC1-lacZ chimera was replaced by a sequence containing a heat shock upstream activating sequence (heat shock element 2) from the 5'-regulatory region of SSA1 was inhibited by excess Ssa1p. The repression of an SSA1 upstream activating sequence by the SSA1 protein indicates that SSA1 self-regulation is at least partially mediated at the transcriptional level. The expression of another transcriptional fusion vector, containing heat shock element 2 and a lesser amount of flanking sequence, is not inhibited when Ssa1p is overexpressed. This suggests the existence of an element, proximal to or overlapping heat shock element 2, that confers sensitivity to the SSA1 protein.

scholarly journals Dual Repression by Fe2+-Fur and Mn2+-MntR of the mntH Gene, Encoding an NRAMP-Like Mn2+ Transporter in Escherichia coli

2001 ◽  
Vol 183 (16) ◽  
pp. 4806-4813 ◽  
Author(s):  
Silke I. Patzer ◽  
Klaus Hantke
Keyword(s):  
Escherichia Coli ◽  
Consensus Sequence ◽  
Transcriptional Regulator ◽  
Binding Region ◽  
Gene Encoding ◽  
Lower Affinity ◽  
E Coli ◽  
Metal Transporter ◽  
Dnase I Footprinting

ABSTRACT The uptake of Mn2+, a cofactor for several enzymes inEscherichia coli, is mediated by MntH, a proton-dependent metal transporter, which also recognizes Fe2+ with lower affinity. MntH belongs to the NRAMP family of eukaryotic Fe2+ and Mn2+ transporters. In E. coli strains with chromosomal mntH-lacZ fusions,mntH was partially repressed by both Mn2+ and Fe2+. Inactivation of fur resulted in the loss of Fe2+-dependent repression of mntHtranscription, demonstrating that Fe2+ repression depends on the global iron regulator Fur. However, these furmutants still showed Mn2+-dependent repression ofmntH. The Mn2+-responsive transcriptional regulator of mntH was identified as the gene product ofo155 (renamed MntR). mntR mutants were impaired in Mn2+ but not Fe2+ repression ofmntH transcription. Binding of purified MntR to themntH operator was manganese dependent. The binding region was localized by DNase I footprinting analysis and covers a nearly perfect palindrome. The Fur binding site, localized within 22 nucleotides of the mntH operator by in vivo operator titration assays, resembles the Fur-box consensus sequence.

Characterization of LgnR, an IclR family transcriptional regulator involved in the regulation of l-gluconate catabolic genes in Paracoccus sp. 43P

Microbiology ◽  
2014 ◽  
Vol 160 (3) ◽  
pp. 623-634 ◽  
Author(s):  
Tetsu Shimizu ◽  
Akira Nakamura
Keyword(s):  
Constitutive Expression ◽  
Transcriptional Regulator ◽  
Pcr Analysis ◽  
Mobility Shift ◽  
Gene Encoding ◽  
Reverse Transcription Pcr ◽  
Dnase I Footprinting ◽  
Catabolic Genes ◽  
Genes Encoding ◽  
Mobility Shift Assay

Five genes encoding enzymes required for l-gluconate catabolism, together with genes encoding components of putative ABC transporters, are located in a cluster in the genome of Paracoccus sp. 43P. A gene encoding a transcriptional regulator in the IclR family, lgnR, is located in front of the cluster in the opposite direction. Reverse transcription PCR analysis indicated that the cluster was transcribed as an operon, termed the lgn operon. Two promoters, P lgnA and P lgnR , are divergently located in the intergenic region, and transcription from these promoters was induced by addition of l-gluconate or d-idonate, a catabolite of l-gluconate. Deletion of lgnR resulted in constitutive expression of lgnA, lgnH and lgnR, indicating that lgnR encodes a repressor protein for the expression of the lgn operon and lgnR itself. Electrophoretic mobility shift assay and DNase I footprinting analyses revealed that recombinant LgnR binds to both P lgnA and P lgnR , indicating that LgnR represses transcription from these promoters by competing with RNA polymerase for binding to these sequences. d-Idonate was identified as a candidate effector molecule for dissociation of LgnR from these promoters. Phylogenetic analysis revealed that LgnR formed a cluster with putative proteins from other genome sequences, which is distinct from those proteins of known regulatory functions, in the IclR family of transcriptional regulators. Additionally, the phylogeny suggests an evolutionary linkage between the l-gluconate catabolic pathway and d-galactonate catabolic pathways distributed in Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria and Actinobacteria.

scholarly journals Molecular Cloning and Transcriptional Regulation of the Aspergillus nidulans xlnD Gene Encoding a β-Xylosidase

1998 ◽  
Vol 64 (4) ◽  
pp. 1412-1419 ◽  
Author(s):  
José A. Pérez-González ◽  
Noël N. M. E. van Peij ◽  
Alja Bezoen ◽  
Andrew P. Maccabe ◽  
Daniel Ramón ◽  
...  
Keyword(s):  
Transcriptional Regulation ◽  
Aspergillus Nidulans ◽  
Binding Sites ◽  
Glucose Repression ◽  
Regulatory Protein ◽  
Regulatory Region ◽  
Northern Analysis ◽  
Transcriptional Level ◽  
Gene Encoding ◽  
Primary Structures

ABSTRACT The xlnD gene encoding the 85-kDa β-xylosidase was cloned from Aspergillus nidulans. The deduced primary structure of the protein exhibits considerable similarity to the primary structures of the Aspergillus niger andTrichoderma reesei β-xylosidases and some similarity to the primary structures of the class 3 β-glucosidases.xlnD is regulated at the transcriptional level; it is induced by xylan and d-xylose and is repressed byd-glucose. Glucose repression is mediated by the product of the creA gene. Although several binding sites for the pH regulatory protein PacC were found in the upstream regulatory region, it was not clear from a Northern analysis whether PacC is involved in transcriptional regulation of xlnD.

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