scholarly journals A complex regulatory element from the yeast gene ENO2 modulates GCR1-dependent transcriptional activation.

1993 ◽  
Vol 13 (4) ◽  
pp. 2623-2633 ◽  
Author(s):  
C E Willett ◽  
C M Gelfman ◽  
M J Holland
Keyword(s):  
Binding Site ◽  
Transcriptional Activation ◽  
Binding Sites ◽  
Regulatory Element ◽  
Enhancer Element ◽  
Cis Acting ◽  
Genes Encoding ◽  
High Level ◽  
Yeast Genes ◽  
Yeast Enolase

The GCR1 gene product is required for maximal transcription of many yeast genes including genes encoding glycolytic enzymes. Transcription of the yeast enolase gene ENO2 is reduced 50-fold in strains carrying a gcr1 null mutation. cis-acting sequences that are sufficient for GCR1-dependent regulation of ENO2 expression were identified by using an enhancerless CYC1 promoter which is not normally dependent on GCR1 for expression. A 60-bp ENO2 sequence that was sufficient to provide high-level, GCR1-dependent transcriptional activation of the CYC1 promoter was identified. This 60-bp element could be subdivided into a 30-bp sequence containing a novel RAP1-binding site and a GCR1-binding site which did not activate CYC1 transcription and a 30-bp sequence containing a novel enhancer element that conferred moderate levels of GCR1-independent transcriptional activation. The 60-bp CGCR1-dependent upstream activator sequence is located immediately downstream from previously mapped overlapping binding sites for the regulatory proteins ABFI and RAP1. Evidence is presented that the overlapping ABFI- and RAP1-binding sites function together with sequences that bind GCR1 and RAP1 to stage transcriptional activation of ENO2 expression.

scholarly journals A complex regulatory element from the yeast gene ENO2 modulates GCR1-dependent transcriptional activation

1993 ◽  
Vol 13 (4) ◽  
pp. 2623-2633
Author(s):  
C E Willett ◽  
C M Gelfman ◽  
M J Holland
Keyword(s):  
Binding Site ◽  
Transcriptional Activation ◽  
Binding Sites ◽  
Regulatory Element ◽  
Enhancer Element ◽  
Cis Acting ◽  
Genes Encoding ◽  
High Level ◽  
Yeast Genes ◽  
Yeast Enolase

The GCR1 gene product is required for maximal transcription of many yeast genes including genes encoding glycolytic enzymes. Transcription of the yeast enolase gene ENO2 is reduced 50-fold in strains carrying a gcr1 null mutation. cis-acting sequences that are sufficient for GCR1-dependent regulation of ENO2 expression were identified by using an enhancerless CYC1 promoter which is not normally dependent on GCR1 for expression. A 60-bp ENO2 sequence that was sufficient to provide high-level, GCR1-dependent transcriptional activation of the CYC1 promoter was identified. This 60-bp element could be subdivided into a 30-bp sequence containing a novel RAP1-binding site and a GCR1-binding site which did not activate CYC1 transcription and a 30-bp sequence containing a novel enhancer element that conferred moderate levels of GCR1-independent transcriptional activation. The 60-bp CGCR1-dependent upstream activator sequence is located immediately downstream from previously mapped overlapping binding sites for the regulatory proteins ABFI and RAP1. Evidence is presented that the overlapping ABFI- and RAP1-binding sites function together with sequences that bind GCR1 and RAP1 to stage transcriptional activation of ENO2 expression.

scholarly journals Vibrio cholerae ToxT Independently Activates the Divergently Transcribed aldA and tagA Genes

2005 ◽  
Vol 187 (23) ◽  
pp. 7890-7900 ◽  
Author(s):  
Jeffrey H. Withey ◽  
Victor J. DiRita
Keyword(s):  
Small Intestine ◽  
Binding Site ◽  
Vibrio Cholerae ◽  
Virulence Factors ◽  
Transcriptional Activation ◽  
Binding Sites ◽  
Inverted Repeat ◽  
Transcriptional Regulators ◽  
Dna Binding Sites ◽  
Genes Encoding

ABSTRACT The Vibrio cholerae ToxT regulon includes the genes encoding cholera toxin (CT) and the toxin-coregulated pilus (TCP), which are the major virulence factors required for causing cholera disease and colonizing the upper small intestine of the host, respectively. The genes encoding CT, ctxAB, and the genes encoding the components of the TCP, tcpA to tcpJ, are organized within operons, upstream of which are DNA binding sites for the transcriptional activator ToxT. ToxT is a member of the large AraC/XylS family of transcriptional regulators and also activates transcription of five other genes whose roles in V. cholerae pathogenesis, if any, are poorly understood. acfA and acfD are divergently transcribed genes required for efficient colonization of the intestine. Transcriptional activation of acfA and acfD requires a pair of central ToxT binding sites in an inverted-repeat configuration for ToxT-directed transcription of both genes. tcpI has an unknown role in pathogenesis. aldA and tagA are divergently transcribed genes that also have unknown roles in pathogenesis. In this study, we map the aldA and tagA promoters and identify the ToxT binding sites upstream of each gene. Our results suggest that two ToxT binding sites in an inverted-repeat configuration are required for ToxT-directed transcription of tagA and that a single ToxT binding site is required for ToxT-directed transcription of aldA. Furthermore, to direct transcription of tagA and aldA, ToxT uses independent binding regions upstream of each gene, in contrast to what we previously found for the divergently transcribed acfA and acfD genes, which share ToxT binding sites between the two genes.

Multiple factors bind the upstream activation sites of the yeast enolase genes ENO1 and ENO2: ABFI protein, like repressor activator protein RAP1, binds cis-acting sequences which modulate repression or activation of transcription

1990 ◽  
Vol 10 (9) ◽  
pp. 4872-4885
Author(s):  
P K Brindle ◽  
J P Holland ◽  
C E Willett ◽  
M A Innis ◽  
M J Holland
Keyword(s):  
Transcriptional Activation ◽  
Binding Sites ◽  
Regulatory Gene ◽  
Regulation Of Gene Expression ◽  
Protein A ◽  
Deletion Mapping ◽  
Autonomously Replicating Sequence ◽  
Cis Acting ◽  
Binding Factor ◽  
Yeast Enolase

Binding sites for three distinct proteins were mapped within the upstream activation sites (UAS) of the yeast enolase genes ENO1 and ENO2. Sequences that overlapped the UAS1 elements of both enolase genes bound a protein which was identified as the product of the RAP1 regulatory gene. Sequences within the UAS2 element of the ENO2 gene bound a second protein which corresponded to the ABFI (autonomously replicating sequence-binding factor) protein. A protein designated EBF1 (enolase-binding factor) bound to sequences which overlapped the UAS2 element in ENO1. There was a good correlation among all of the factor-binding sites and the location of sequences required for UAS activity identified by deletion mapping analysis. This observation suggested that the three factors play a role in transcriptional activation of the enolase genes. UAS elements which bound the RAP1 protein or the ABFI protein modulated glucose-dependent induction of ENO1 and ENO2 expression. The ABFI-binding site in ENO2 overlapped sequences required for UAS2 activity in wild-type strains and for repression of ENO2 expression in strains carrying a null mutation in the positive regulatory gene GCR1. These latter results showed that the ABFI protein, like the RAP1 protein, bound sequences required for positive as well as negative regulation of gene expression. These observations strongly suggest that the biological functions of the RAP1 and ABFI proteins are similar.

scholarly journals Interactions of Sox10 and Egr2 in myelin gene regulation

2007 ◽  
Vol 3 (4) ◽  
pp. 377-387 ◽  
Author(s):  
Erin A. Jones ◽  
Sung-Wook Jang ◽  
Gennifer M. Mager ◽  
Li-Wei Chang ◽  
Rajini Srinivasan ◽  
...  
Keyword(s):  
Binding Sites ◽  
Basic Protein ◽  
Developmental Regulation ◽  
Regulatory Element ◽  
Enhancer Element ◽  
First Intron ◽  
Genes Encoding ◽  
Myelin Associated Glycoprotein ◽  
Protein Zero ◽  
Myelin Gene

AbstractMyelination in the PNS is accompanied by a large induction of the myelin protein zero (Mpz) gene to produce the most abundant component in peripheral myelin. Analyses of knockout mice have shown that the EGR2/Krox20 and SOX10 transcription factors are required for Mpz expression. Our recent work has shown that the dominant EGR2 mutations associated with human peripheral neuropathies cause disruption of EGR2/SOX10 synergy at specific sites, including a conserved enhancer element in the first intron of the Mpz gene. Further investigation of Egr2/Sox10 interactions reveals that activation of the Mpz intron element by Egr2 requires both Sox10-binding sites. In addition, both Egr1 and Egr3 cooperate with Sox10 to activate this element, which indicates that this capacity is conserved among Egr family members. Finally, a conserved composite structure of Egr2/Sox10-binding sites in the genes encoding Mpz, myelin-associated glycoprotein and myelin basic protein genes was used to screen for similar modules in other myelin genes, revealing a potential regulatory element in the periaxin gene. Overall, these results elucidate a working model for developmental regulation of Mpz expression, several facets of which extend to regulation of other peripheral myelin genes.

scholarly journals Multiple factors bind the upstream activation sites of the yeast enolase genes ENO1 and ENO2: ABFI protein, like repressor activator protein RAP1, binds cis-acting sequences which modulate repression or activation of transcription.

1990 ◽  
Vol 10 (9) ◽  
pp. 4872-4885 ◽  
Author(s):  
P K Brindle ◽  
J P Holland ◽  
C E Willett ◽  
M A Innis ◽  
M J Holland
Keyword(s):  
Transcriptional Activation ◽  
Binding Sites ◽  
Regulatory Gene ◽  
Regulation Of Gene Expression ◽  
Protein A ◽  
Deletion Mapping ◽  
Autonomously Replicating Sequence ◽  
Cis Acting ◽  
Binding Factor ◽  
Yeast Enolase

Binding sites for three distinct proteins were mapped within the upstream activation sites (UAS) of the yeast enolase genes ENO1 and ENO2. Sequences that overlapped the UAS1 elements of both enolase genes bound a protein which was identified as the product of the RAP1 regulatory gene. Sequences within the UAS2 element of the ENO2 gene bound a second protein which corresponded to the ABFI (autonomously replicating sequence-binding factor) protein. A protein designated EBF1 (enolase-binding factor) bound to sequences which overlapped the UAS2 element in ENO1. There was a good correlation among all of the factor-binding sites and the location of sequences required for UAS activity identified by deletion mapping analysis. This observation suggested that the three factors play a role in transcriptional activation of the enolase genes. UAS elements which bound the RAP1 protein or the ABFI protein modulated glucose-dependent induction of ENO1 and ENO2 expression. The ABFI-binding site in ENO2 overlapped sequences required for UAS2 activity in wild-type strains and for repression of ENO2 expression in strains carrying a null mutation in the positive regulatory gene GCR1. These latter results showed that the ABFI protein, like the RAP1 protein, bound sequences required for positive as well as negative regulation of gene expression. These observations strongly suggest that the biological functions of the RAP1 and ABFI proteins are similar.

Role of AP1/NFE2 binding sites in endogenous α-globin gene transcription

Blood ◽  
2003 ◽  
Vol 102 (12) ◽  
pp. 4223-4228 ◽  
Author(s):  
Melanie R. Loyd ◽  
Yasuhiro Okamoto ◽  
Mindy S. Randall ◽  
Paul A. Ney
Keyword(s):  
Gene Transcription ◽  
Binding Sites ◽  
Regulatory Element ◽  
Globin Gene ◽  
Gene Activation ◽  
Loxp Site ◽  
Regulatory Sequences ◽  
Cis Acting ◽  
High Level

Abstract High-level α-globin expression depends on cis-acting regulatory sequences located far upstream of the α-globin cluster. Sequences that contain the α-globin positive regulatory element (PRE) activate α-globin expression in transgenic mice. The α-globin PRE contains a pair of composite binding sites for the transcription factors activating protein 1 and nuclear factor erythroid 2 (AP1/NFE2). To determine the role of these binding sites in α-globin gene transcription, we mutated the AP1/NFE2 sites in the α-globin PRE in mice. We replaced the AP1/NFE2 sites with a neomycin resistance gene (neo) that is flanked by LoxP sites (floxed). Mice with this mutation exhibited increased embryonic death and α-thalassemia intermedia. Next, we removed the neo gene by Cre-mediated recombination, leaving a single LoxP site in place of the AP1/NFE2 sites. These mice were phenotypically normal. However, α-globin expression, measured by allele-specific RNA polymerase chain reaction (PCR), was decreased 25%. We examined the role of the hematopoietic-restricted transcription factor p45Nfe2 in activating expression through these sites and found that it is not required. Thus, we have demonstrated that AP1/NFE2 binding sites in the murine α-globin PRE contribute to long-range α-globin gene activation. The proteins that mediate this effect remain to be determined. (Blood. 2003;102:4223-4228)

scholarly journals Sequences Just Upstream of the Simian Immunodeficiency Virus Core Enhancer Allow Efficient Replication in the Absence of NF-κB and Sp1 Binding Elements

1998 ◽  
Vol 72 (7) ◽  
pp. 5589-5598 ◽  
Author(s):  
Stefan Pöhlmann ◽  
Stefan Flöss ◽  
Petr O. Ilyinskii ◽  
Thomas Stamminger ◽  
Frank Kirchhoff
Keyword(s):  
Viral Replication ◽  
Binding Site ◽  
Simian Immunodeficiency Virus ◽  
Binding Sites ◽  
Regulatory Element ◽  
Virus Production ◽  
Enhancer Element ◽  
Immunodeficiency Virus ◽  
Efficient Replication

ABSTRACT Large deletions of the upstream U3 sequences in the long terminal repeats (LTRs) of human immunodeficiency virus and simian immunodeficiency virus (SIV) accumulate in vivo in the absence of an intact nef gene. In the SIV U3 region, about 65 bp just upstream of the single NF-κB binding site always remained intact, and some evidence for a novel enhancer element in this region exists. We analyzed the transcriptional and replicative capacities of SIVmac239 mutants containing deletions or mutations in these upstream U3 sequences and/or the NF-κB and Sp1 binding sites. Even in the absence of 400 bp of upstream U3 sequences, the NF-κB site and all four Sp1 binding sites, the SIV promoter maintained about 15% of the wild-type LTR activity and was fully responsive to Tat activation in transient reporter assays. The effects of these deletions on virus production after transfection of COS-1 cells with full-length proviral constructs were much greater. Deletion of the upstream U3 sequences had no significant influence on viral replication when either the single NF-κB site or the Sp1 binding sites were intact. In contrast, the 26 bp of sequence located immediately upstream of the NF-κB site was essential for efficient replication when all core enhancer elements were deleted. A purine-rich site in this region binds specifically to the transcription factor Elf-1, a member of the etsproto-oncogene-encoded family. Our results indicate a high degree of functional redundancy in the SIVmac U3 region. Furthermore, we defined a novel regulatory element located immediately upstream of the NF-κB binding site that allows efficient viral replication in the absence of the entire core enhancer region.

scholarly journals Genes encoding components of the olfactory signal transduction cascade contain a DNA binding site that may direct neuronal expression.

1993 ◽  
Vol 13 (9) ◽  
pp. 5805-5813 ◽  
Author(s):  
M M Wang ◽  
R Y Tsai ◽  
K A Schrader ◽  
R R Reed
Keyword(s):  
Signal Transduction ◽  
Dna Binding ◽  
Binding Site ◽  
Binding Sites ◽  
Consensus Sequence ◽  
Initiation Site ◽  
Olfactory Neuron ◽  
Promoter Regions ◽  
Transcriptional Initiation ◽  
Genes Encoding

Genes which mediate odorant signal transduction are expressed at high levels in neurons of the olfactory epithelium. The molecular mechanism governing the restricted expression of these genes likely involves tissue-specific DNA binding proteins which coordinately activate transcription through sequence-specific interactions with olfactory promoter regions. We have identified binding sites for the olfactory neuron-specific transcription factor, Olf-1, in the sequences surrounding the transcriptional initiation site of five olfactory neuron-specific genes. The Olf-1 binding sites described define the consensus sequence YTCCCYRGGGAR. In addition, we have identified a second binding site, the U site, in the olfactory cyclic nucleotide gated channel and type III cyclase promoters, which binds factors present in all tissue examined. These experiments support a model in which expression of Olf-1 in the sensory neurons coordinately activates a set of olfactory neuron-specific genes. Furthermore, expression of a subset of these genes may be modulated by additional binding factors.

DNA-binding and transcriptional regulatory properties of hepatic leukemia factor (HLF) and the t(17;19) acute lymphoblastic leukemia chimera E2A-HLF

1994 ◽  
Vol 14 (9) ◽  
pp. 5986-5996
Author(s):  
S P Hunger ◽  
R Brown ◽  
M L Cleary
Keyword(s):  
Dna Binding ◽  
Binding Site ◽  
Dna Sequences ◽  
Transcriptional Activation ◽  
Binding Sites ◽  
Lymphoblastic Leukemia ◽  
Consensus Sequence ◽  
Reporter Genes ◽  
Wild Type ◽  
Basic Leucine Zipper

The t(17;19) translocation in acute lymphoblastic leukemias results in creation of E2A-hepatic leukemia factor (HLF) chimeric proteins that contain the DNA-binding and protein dimerization domains of the basic leucine zipper (bZIP) protein HLF fused to a portion of E2A proteins with transcriptional activation properties. An in vitro binding site selection procedure was used to determine DNA sequences preferentially bound by wild-type HLF and chimeric E2A-HLF proteins isolated from various t(17;19)-bearing leukemias. All were found to selectively bind the consensus sequence 5'-GTTACGTAAT-3' with high affinity. Wild-type and chimeric HLF proteins also bound closely related sites identified previously for bZIP proteins of both the proline- and acidic amino acid-rich (PAR) and C/EBP subfamilies; however, E2A-HLF proteins were significantly less tolerant of certain deviations from the HLF consensus binding site. These differences were directly attributable to loss of an HLF ancillary DNA-binding domain in all E2A-HLF chimeras and were further exacerbated by a zipper mutation in one isolate. Both wild-type and chimeric HLF proteins displayed transcriptional activator properties in lymphoid and nonlymphoid cells on reporter genes containing HLF or C/EBP consensus binding sites. But on reporter genes with nonoptimal binding sites, their transcriptional properties diverged and E2A-HLF competitively inhibited activation by wild-type PAR proteins. These findings establish a spectrum of binding site-specific transcriptional properties for E2A-HLF which may preferentially activate expression of select subordinate genes as a homodimer and potentially antagonize expression of others through heteromeric interactions.

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