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Article ; Online: Retraction notice to "TGF-β1 regulation of human AT1 receptor mRNA splice variants harboring exon 2".

Martin, Mickey M / Buckenberger, Jessica A / Knoell, Daren L / Strauch, Arthur R / Elton, Terry S

2016 Volume 434, Page(s) 288

Language	English
Publishing date	2016-10-15
Publishing country	Ireland
Document type	Published Erratum ; Retraction of Publication
ZDB-ID	187438-x
ISSN	1872-8057 ; 0303-7207
ISSN (online)	1872-8057
ISSN	0303-7207
DOI	10.1016/j.mce.2016.08.003
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Article: Building better blood vessels: new insight on the molecular control of arteriogenesis.

Strauch, Arthur R

Cardiovascular research

2003 Volume 59, Issue 3, Page(s) 532–533

MeSH term(s)	Animals ; Arterial Occlusive Diseases/immunology ; Arterial Occlusive Diseases/metabolism ; Arterial Occlusive Diseases/pathology ; Arteries/immunology ; Arteries/metabolism ; Arteries/pathology ; Carbonic Anhydrases ; Collateral Circulation/physiology ; DNA-Binding Proteins/metabolism ; Early Growth Response Protein 1 ; Endothelial Cells/immunology ; Endothelial Cells/metabolism ; Endothelial Cells/pathology ; Gene Expression ; Humans ; Immediate-Early Proteins ; MAP Kinase Signaling System ; Mice ; Neoplasms/blood supply ; Neoplasms/immunology ; Neoplasms/pathology ; Neovascularization, Pathologic/pathology ; Nerve Tissue Proteins/genetics ; Stress, Mechanical ; Transcription Factors/metabolism ; Transforming Growth Factor beta/metabolism ; Tunica Intima/immunology ; Tunica Intima/metabolism ; Tunica Intima/pathology
Chemical Substances	DNA-Binding Proteins ; EGR1 protein, human ; Early Growth Response Protein 1 ; Egr1 protein, mouse ; Immediate-Early Proteins ; Nerve Tissue Proteins ; Transcription Factors ; Transforming Growth Factor beta ; Carbonic Anhydrases (EC 4.2.1.1)
Language	English
Publishing date	2003-09-01
Publishing country	England
Document type	Comment ; Editorial
ZDB-ID	80340-6
ISSN	1755-3245 ; 0008-6363
ISSN (online)	1755-3245
ISSN	0008-6363
DOI	10.1016/s0008-6363(03)00504-2
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Article ; Online: Transforming growth factor beta1-mediated activation of the smooth muscle alpha-actin gene in human pulmonary myofibroblasts is inhibited by tumor necrosis factor-alpha via mitogen-activated protein kinase kinase 1-dependent induction of the Egr-1 transcriptional repressor.

Liu, Xiaoying / Kelm, Robert J / Strauch, Arthur R

Molecular biology of the cell

2009 Volume 20, Issue 8, Page(s) 2174–2185

Abstract: Transforming growth factor (TGF) beta1 is a mediator of myofibroblast differentiation in healing wounds in which it activates transcription of the smooth muscle alpha-actin (SMalphaA) gene via dynamic interplay of nuclear activators and repressors. ... ...

Abstract	Transforming growth factor (TGF) beta1 is a mediator of myofibroblast differentiation in healing wounds in which it activates transcription of the smooth muscle alpha-actin (SMalphaA) gene via dynamic interplay of nuclear activators and repressors. Targeting components of TGFbeta1 signaling may be an effective strategy for controlling myofibroblasts in chronic fibrotic diseases. We examined the ability of proinflammatory tumor necrosis factor (TNF)-alpha to antagonize TGFbeta1-mediated human pulmonary myofibroblast differentiation. TNF-alpha abrogated TGFbeta1-induced SMalphaA gene expression at the level of transcription without disrupting phosphorylation of regulatory Smads. Intact mitogen-activated protein kinase kinase (Mek)-extracellular signal-regulated kinase (Erk) kinase signaling was required for myofibroblast repression by TNF-alpha via induction of the early growth response factor-1 (Egr-1) DNA-binding protein. Egr-1 bound to the GC-rich SPUR activation element in the SMalphaA promoter and potently suppressed Smad3- and TGFbeta1-mediated transcription. Reduction in Smad binding to the SMalphaA promoter in TNF-alpha-treated myofibroblasts was accompanied by an increase in Egr-1 and YB-1 repressor binding, suggesting that the molecular mechanism underlying repression may involve competitive interplay between Egr-1, YB-1, and Smads. The ability of TNF-alpha to attenuate myofibroblast differentiation via modulation of a Mek1/Erk/Egr-1 regulatory axis may be useful in designing new therapeutic targets to offset destructive tissue remodeling in chronic fibrotic disease.
MeSH term(s)	Actins/genetics ; Early Growth Response Protein 1/metabolism ; Extracellular Signal-Regulated MAP Kinases/metabolism ; Fibroblasts/drug effects ; Fibroblasts/enzymology ; Gene Expression Regulation/drug effects ; Humans ; Lung/cytology ; MAP Kinase Kinase 1/metabolism ; Models, Genetic ; Muscle, Smooth/metabolism ; Phosphorylation/drug effects ; Promoter Regions, Genetic/genetics ; Protein Binding/drug effects ; Repressor Proteins/metabolism ; Smad2 Protein/metabolism ; Smad7 Protein/metabolism ; Transcription, Genetic/drug effects ; Transforming Growth Factor beta1/pharmacology ; Tumor Necrosis Factor-alpha/pharmacology ; Y-Box-Binding Protein 1/metabolism
Chemical Substances	Actins ; EGR1 protein, human ; Early Growth Response Protein 1 ; Repressor Proteins ; SMAD2 protein, human ; SMAD7 protein, human ; Smad2 Protein ; Smad7 Protein ; Transforming Growth Factor beta1 ; Tumor Necrosis Factor-alpha ; Y-Box-Binding Protein 1 ; Extracellular Signal-Regulated MAP Kinases (EC 2.7.11.24) ; MAP Kinase Kinase 1 (EC 2.7.12.2)
Language	English
Publishing date	2009-03-04
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	1098979-1
ISSN	1939-4586 ; 1059-1524
ISSN (online)	1939-4586
ISSN	1059-1524
DOI	10.1091/mbc.E08-10-0994
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Zs.A 2853: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
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Article ; Online: Polyclonal F(ab')

Cunha, Luis Eduardo R / Stolet, Adilson A / Strauch, Marcelo A / Pereira, Victor A R / Dumard, Carlos H / Gomes, Andre M O / Monteiro, Fábio L / Higa, Luiza M / Souza, Patrícia N C / Fonseca, Juliana G / Pontes, Francisco E / Meirelles, Leonardo G R / Albuquerque, José W M / Sacramento, Carolina Q / Fintelman-Rodrigues, Natalia / Lima, Tulio M / Alvim, Renata G F / Marsili, Federico F / Caldeira, Marcella Moreira /

Zingali, Russolina B / de Oliveira, Guilherme A P / Souza, Thiago M L / Silva, Alexandre S / Muller, Rodrigo / Rodrigues, Daniela Del Rosário Flores / Jesus da Costa, Luciana / Alves, Arthur Daniel R / Pinto, Marcelo Alves / Oliveira, Andréa C / Guedes, Herbert L M / Tanuri, Amilcar / Castilho, Leda R / Silva, Jerson L

iScience

2021 Volume 24, Issue 11, Page(s) 103315

Abstract: We used the recombinant trimeric spike (S) glycoprotein in the prefusion conformation to immunize horses for the production of hyperimmune globulins against SARS-CoV-2. Serum antibody titers measured by ELISA were above 1: ... ...

Abstract	We used the recombinant trimeric spike (S) glycoprotein in the prefusion conformation to immunize horses for the production of hyperimmune globulins against SARS-CoV-2. Serum antibody titers measured by ELISA were above 1:10
Language	English
Publishing date	2021-10-23
Publishing country	United States
Document type	Journal Article
ISSN	2589-0042
ISSN (online)	2589-0042
DOI	10.1016/j.isci.2021.103315
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Article ; Online: Y-box binding protein-1 implicated in translational control of fetal myocardial gene expression after cardiac transplant.

David, Jason J / Subramanian, Sukanya V / Zhang, Aiwen / Willis, William L / Kelm, Robert J / Leier, Carl V / Strauch, Arthur R

Experimental biology and medicine (Maywood, N.J.)

2012 Volume 237, Issue 5, Page(s) 593–607

Abstract: Peri-transplant surgical trauma and ischemia/reperfusion injury in accepted murine heterotopic heart grafts has been associated with myofibroblast differentiation, cardiac fibrosis and biomechanical-stress activation of the fetal myocardial smooth muscle ...

Abstract	Peri-transplant surgical trauma and ischemia/reperfusion injury in accepted murine heterotopic heart grafts has been associated with myofibroblast differentiation, cardiac fibrosis and biomechanical-stress activation of the fetal myocardial smooth muscle α-actin (SMαA) gene. The wound-healing agonists, transforming growth factor β1 and thrombin, are known to coordinate SMαA mRNA transcription and translation in activated myofibroblasts by altering the subcellular localization and mRNA-binding affinity of the Y-box binding protein-1 (YB-1) cold-shock domain (CSD) protein that governs a variety of cellular responses to metabolic stress. YB-1 accumulated in polyribosome-enriched regions of the sarcoplasm proximal to cardiac intercalated discs in accepted heart grafts. YB-1 binding to a purine-rich motif in exon 3 of SMαA mRNA that regulates translational efficiency increased substantially in perfusion-isolated, rod-shaped adult rat cardiomyocytes during phenotypic de-differentiation in the presence of serum-derived growth factors. Cardiomyocyte de-differentiation was accompanied by the loss of a 60 kDa YB-1 variant that was highly expressed in both adult myocardium and freshly isolated myocytes and replacement with the 50 kDa form of YB-1 (p50) typically expressed in myofibroblasts that demonstrated sequence-specific interaction with SMαA mRNA. Accumulation of p50 YB-1 in reprogrammed, de-differentiated myocytes was associated with a 10-fold increase in SMαA protein expression. Endomyocardial biopsies collected from patients up to 14 years after heart transplant showed variable yet coordinately elevated expression of SMαA and p50 YB-1 protein and demonstrable p50 YB-1:SMαA mRNA interaction. The p60 YB-1 variant in human heart graft samples, but neither mouse p60 nor mouse or human p50, reacted with an antibody specific for the phosphoserine 102 modification in the YB-1 CSD. Modulation of YB-1 subcellular compartmentalization and mRNA-binding activity may be linked with reprogramming of contractile protein gene expression in ventricular cardiomyocytes that could contribute to maladaptive remodeling in accepted, long-term heart grafts.
MeSH term(s)	Actins/genetics ; Actins/metabolism ; Animals ; Animals, Newborn ; Electrophoretic Mobility Shift Assay ; Gene Expression ; Heart Transplantation ; Humans ; Mice ; Mice, Inbred C57BL ; Mice, Inbred DBA ; Muscle, Smooth, Vascular/metabolism ; Myocardial Reperfusion Injury/metabolism ; Myocardium/metabolism ; Myocytes, Cardiac/metabolism ; Myofibroblasts/metabolism ; Promoter Regions, Genetic ; Protein Biosynthesis ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Rats ; Rats, Sprague-Dawley ; Transcription, Genetic ; Transplantation, Heterotopic ; Wound Healing ; Y-Box-Binding Protein 1/genetics ; Y-Box-Binding Protein 1/metabolism
Chemical Substances	Actins ; RNA, Messenger ; Y-Box-Binding Protein 1 ; YBX1 protein, human ; alpha-smooth muscle actin, mouse
Language	English
Publishing date	2012-05
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	4015-0
ISSN	1535-3699 ; 1525-1373 ; 0037-9727
ISSN (online)	1535-3699 ; 1525-1373
ISSN	0037-9727
DOI	10.1258/ebm.2012.011137
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Article ; Online: Polyclonal F(ab’)2 fragments of equine antibodies raised against the spike protein neutralize SARS-CoV-2 variants with high potency

Luis Eduardo R. Cunha / Adilson A. Stolet / Marcelo A. Strauch / Victor A.R. Pereira / Carlos H. Dumard / Andre M.O. Gomes / Fábio L. Monteiro / Luiza M. Higa / Patrícia N.C. Souza / Juliana G. Fonseca / Francisco E. Pontes / Leonardo G.R. Meirelles / José W.M. Albuquerque / Carolina Q. Sacramento / Natalia Fintelman-Rodrigues / Tulio M. Lima / Renata G.F. Alvim / Federico F. Marsili / Marcella Moreira Caldeira /

Russolina B. Zingali / Guilherme A.P. de Oliveira / Thiago M.L. Souza / Alexandre S. Silva / Rodrigo Muller / Daniela del Rosário Flores Rodrigues / Luciana Jesus da Costa / Arthur Daniel R. Alves / Marcelo Alves Pinto / Andréa C. Oliveira / Herbert L.M. Guedes / Amilcar Tanuri / Leda R. Castilho / Jerson L. Silva

iScience, Vol 24, Iss 11, Pp 103315- (2021)

2021

Abstract: Summary: We used the recombinant trimeric spike (S) glycoprotein in the prefusion conformation to immunize horses for the production of hyperimmune globulins against SARS-CoV-2. Serum antibody titers measured by ELISA were above 1:106, and the ... ...

Abstract	Summary: We used the recombinant trimeric spike (S) glycoprotein in the prefusion conformation to immunize horses for the production of hyperimmune globulins against SARS-CoV-2. Serum antibody titers measured by ELISA were above 1:106, and the neutralizing antibody titer against authentic virus (WT) was 1:14,604 (average PRNT90). Plasma from immunized animals was pepsin digested to remove the Fc portion and purified, yielding an F(ab’)2 preparation with PRNT90 titers 150-fold higher than the neutralizing titers in human convalescent plasma. Challenge studies were carried out in hamsters and showed the in vivo ability of equine F(ab’)2 to reduce viral load in the pulmonary tissues and significant clinical improvement determined by weight gain. The neutralization curve by F(ab’)2 was similar against the WT and P.2 variants, but displaced to higher concentrations by 0.39 log units against the P.1 (Gamma) variant. These results support the possibility of using equine F(ab’)2 preparation for the clinical treatment of COVID patients.
Keywords	Immunology ; Equine immunology ; Biological sciences ; Science ; Q
Subject code	630
Language	English
Publishing date	2021-11-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
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Article: Structure-function analysis of mouse Pur beta II. Conformation altering mutations disrupt single-stranded DNA and protein interactions crucial to smooth muscle alpha-actin gene repression.

Knapp, Anna M / Ramsey, Jon E / Wang, Shu-Xia / Strauch, Arthur R / Kelm, Robert J

The Journal of biological chemistry

2007 Volume 282, Issue 49, Page(s) 35899–35909

Abstract: Previous studies from our laboratories have implicated two members of the Pur family of single-stranded DNA/RNA-binding proteins, Pur alpha and Pur beta, in transcriptional repression of the smooth muscle alpha-actin gene in vascular cell types. Although ...

Abstract	Previous studies from our laboratories have implicated two members of the Pur family of single-stranded DNA/RNA-binding proteins, Pur alpha and Pur beta, in transcriptional repression of the smooth muscle alpha-actin gene in vascular cell types. Although Pur alpha and Pur beta share substantial sequence homology and nucleic acid binding properties, genomic promoter and cis-element occupancy studies reported herein suggest that Pur beta is the dominant factor in gene regulation. To dissect the molecular basis of Pur beta repressor activity, site-directed mutagenesis was used to map amino acids critical to the physical and functional interaction of Pur beta with the smooth muscle alpha-actin promoter. Of all the various acidic, basic, and aromatic residues studied, mutation of positionally conserved arginines in the class I or class II repeat modules significantly attenuated Pur beta repressor activity in transfected vascular smooth muscle cells and fibroblasts. DNA binding and protein-protein interaction assays were conducted with purified recombinant Pur beta and selected mutants to reveal the physical basis for loss-of-function. Mutants R57E, R57E/R96E, and R57A/R96A each exhibited reduced single-stranded DNA binding affinity for an essential promoter element and diminished interaction with corepressor YB-1/MSY1. Structural analyses of the R57A/R96A and R57E/R96E double mutants in comparison to the wild-type Pur beta homodimer revealed aberrant self-association into higher order oligomeric complexes, which correlated with decreased alpha-helical content and defective DNA and protein binding in vitro. These findings point to a previously unrecognized structural role for certain core arginine residues in forming a conformationally stable Pur beta protein capable of physical interactions necessary for smooth muscle alpha-actin gene repression.
MeSH term(s)	Actins/biosynthesis ; Actins/genetics ; Amino Acid Substitution ; Animals ; Cell Line ; DNA, Single-Stranded/genetics ; DNA, Single-Stranded/metabolism ; DNA-Binding Proteins/genetics ; DNA-Binding Proteins/metabolism ; Fibroblasts/cytology ; Fibroblasts/metabolism ; Gene Silencing ; Mice ; Muscle, Smooth, Vascular/cytology ; Muscle, Smooth, Vascular/metabolism ; Mutagenesis, Site-Directed ; Mutation, Missense ; Myocytes, Smooth Muscle/cytology ; Myocytes, Smooth Muscle/metabolism ; Nerve Tissue Proteins/genetics ; Nerve Tissue Proteins/metabolism ; Protein Binding/genetics ; Protein Structure, Quaternary ; Protein Structure, Secondary ; Rats ; Recombinant Proteins/genetics ; Recombinant Proteins/metabolism ; Repressor Proteins/genetics ; Repressor Proteins/metabolism ; Response Elements/physiology ; Sequence Homology, Amino Acid ; Structure-Activity Relationship
Chemical Substances	Actins ; DNA, Single-Stranded ; DNA-Binding Proteins ; Nerve Tissue Proteins ; Nsep1 protein, mouse ; Pura protein, mouse ; Purb protein, mouse ; Recombinant Proteins ; Repressor Proteins
Language	English
Publishing date	2007-09-28
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2997-x
ISSN	1083-351X ; 0021-9258
ISSN (online)	1083-351X
ISSN	0021-9258
DOI	10.1074/jbc.M706617200
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Article: TGF-beta(1) regulation of human AT(1) receptor mRNA splice variants harboring exon 2.

Martin, Mickey M / Buckenberger, Jessica A / Knoell, Daren L / Strauch, Arthur R / Elton, Terry S

publication RETRACTED

Molecular and cellular endocrinology

2006 Volume 249, Issue 1-2, Page(s) 21–31

Abstract: At least four alternatively spliced mRNAs can be synthesized from the human AT(1)R (hAT(1)R) gene ... variants harboring exon 2 accounts for at least 30% of all the hAT(1)R mRNA transcripts expressed ... we hypothesized that these AUGs would inhibit the translation of the downstream hAT(1)R protein ORF harbored ...

Abstract	At least four alternatively spliced mRNAs can be synthesized from the human AT(1)R (hAT(1)R) gene that differ only in the inclusion or exclusion of exon 2 and/or 3. RT-PCR experiments demonstrate that splice variants harboring exon 2 accounts for at least 30% of all the hAT(1)R mRNA transcripts expressed in the human tissues investigated. Since exon 2 contains two upstream AUGs or open reading frames (uORFs), we hypothesized that these AUGs would inhibit the translation of the downstream hAT(1)R protein ORF harbored in exon 4. This study demonstrates that the inclusion of exon 2 in hAT(1)R mRNA transcripts dramatically reduces hAT(1)R protein levels (nine-fold) and significantly attenuates Ang II responsiveness ( approximately four-fold). Interestingly, only when both AUGs were mutated in combination were the hAT(1)R density and Ang II signaling levels comparable with those values obtained using mRNA splice variants that did not include exon 2. This observation is consistent with a model where the majority of the ribosomes likely translate uORF#1 and are then unable to reinitiate at the downstream hAT(1)R ORF, in part due to the presence of AUG#2 and to the short intercistronic spacing. Importantly, TGF-beta(1) treatment (4ng/ml for 4h) of fibroblasts up-regulated hAT(1)R mRNA splice variants, which harbored exon 2, six-fold. Since AT(1)R activation is closely associated with cardiovascular disease, the inclusion of exon 2 by alternative splicing represents a novel mechanism to reduce the overall production of the hAT(1)R protein and possibly limit the potential pathological effects of AT(1)R activation.
MeSH term(s)	Alternative Splicing ; Base Sequence ; Codon, Initiator/physiology ; Exons ; Humans ; Molecular Sequence Data ; RNA, Messenger/metabolism ; Receptor, Angiotensin, Type 1/biosynthesis ; Receptor, Angiotensin, Type 1/genetics ; Sequence Alignment ; Transforming Growth Factor beta/pharmacology ; Transforming Growth Factor beta/physiology ; Up-Regulation
Chemical Substances	Codon, Initiator ; RNA, Messenger ; Receptor, Angiotensin, Type 1 ; Transforming Growth Factor beta
Language	English
Publishing date	2006--25
Publishing country	Ireland
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Retracted Publication
ZDB-ID	187438-x
ISSN	1872-8057 ; 0303-7207
ISSN (online)	1872-8057
ISSN	0303-7207
DOI	10.1016/j.mce.2006.01.009
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Article: Structure/function analysis of mouse Purbeta, a single-stranded DNA-binding repressor of vascular smooth muscle alpha-actin gene transcription.

Kelm, Robert J / Wang, Shu-Xia / Polikandriotis, John A / Strauch, Arthur R

The Journal of biological chemistry

2003 Volume 278, Issue 40, Page(s) 38749–38757

Abstract: Plasticity of smooth muscle alpha-actin gene expression in fibroblasts and vascular smooth muscle cells is mediated by opposing effects of transcriptional activators and repressors. Among these factors, three single-stranded DNA-binding proteins, ... ...

Abstract	Plasticity of smooth muscle alpha-actin gene expression in fibroblasts and vascular smooth muscle cells is mediated by opposing effects of transcriptional activators and repressors. Among these factors, three single-stranded DNA-binding proteins, Puralpha, Purbeta, and MSY1, have been implicated as coregulators of a cryptic 5'-enhancer module. In this study, a molecular analysis of Purbeta, the least well characterized member of this group, was conducted. Southwestern and Northwestern blotting of purified Purbeta deletion mutants using smooth muscle alpha-actin-derived probes mapped the minimal single-stranded DNA/RNA-binding domain to a conserved region spanning amino acids 37-263. Quantitative binding assays indicated that the relative affinity and specificity of Purbeta for single-stranded DNA were influenced by purine/pyrimidine content; by non-conserved regions outside amino acids 37-263; and by cell-derived proteins, specifically MSY1. When overexpressed in A7r5 vascular smooth muscle cells, Purbeta (but not Puralpha) inhibited transcription of a smooth muscle-specific mouse alpha-actin promoter transgene. Structural domains required for Purbeta repressor activity included the minimal DNA-binding region and a C-terminal domain required for stabilizing high affinity protein and nucleic acid interactions. Purbeta inhibitory activity in transfected A7r5 cells was potentiated by MSY1, but antagonized by serum response factor, reinforcing the idea that interplay among activators and repressors may account for phenotypic changes in smooth muscle alpha-actin-expressing cell types.
MeSH term(s)	Actins/biosynthesis ; Actins/genetics ; Animals ; Blotting, Northern ; Blotting, Southern ; Blotting, Western ; Cell Line ; DNA/metabolism ; DNA, Complementary/metabolism ; DNA, Single-Stranded/metabolism ; DNA-Binding Proteins/chemistry ; DNA-Binding Proteins/metabolism ; DNA-Binding Proteins/physiology ; Dose-Response Relationship, Drug ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli/metabolism ; Gene Deletion ; Genes, Reporter ; Immunoblotting ; Mice ; Muscle, Smooth, Vascular/metabolism ; Mutation ; Oligonucleotides/chemistry ; Phenotype ; Plasmids/metabolism ; Protein Binding ; Protein Structure, Tertiary ; Rats ; Recombinant Proteins/metabolism ; Serum Response Factor/metabolism ; Structure-Activity Relationship ; Time Factors ; Transcription, Genetic ; Transfection ; Transgenes
Chemical Substances	Actins ; DNA, Complementary ; DNA, Single-Stranded ; DNA-Binding Proteins ; Nsep1 protein, mouse ; Oligonucleotides ; Purb protein, mouse ; Recombinant Proteins ; Serum Response Factor ; DNA (9007-49-2)
Language	English
Publishing date	2003-07-21
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, P.H.S.
ZDB-ID	2997-x
ISSN	1083-351X ; 0021-9258
ISSN (online)	1083-351X
ISSN	0021-9258
DOI	10.1074/jbc.M306163200
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Article: Cryptic MCAT enhancer regulation in fibroblasts and smooth muscle cells. Suppression of TEF-1 mediated activation by the single-stranded DNA-binding proteins, Pur alpha, Pur beta, and MSY1.

Carlini, Leslie E / Getz, Michael J / Strauch, Arthur R / Kelm, Robert J

The Journal of biological chemistry

2001 Volume 277, Issue 10, Page(s) 8682–8692

Abstract: An asymmetric polypurine-polypyrimidine cis-element located in the 5' region of the mouse vascular smooth muscle alpha-actin gene serves as a binding site for multiple proteins with specific affinity for either single- or double-stranded DNA. Here, we ... ...

Abstract	An asymmetric polypurine-polypyrimidine cis-element located in the 5' region of the mouse vascular smooth muscle alpha-actin gene serves as a binding site for multiple proteins with specific affinity for either single- or double-stranded DNA. Here, we test the hypothesis that single-stranded DNA-binding proteins are responsible for preventing a cryptic MCAT enhancer centered within this element from cooperating with a nearby serum response factor-interacting CArG motif to trans-activate the minimal promoter in fibroblasts and smooth muscle cells. DNA binding studies revealed that the core MCAT sequence mediates binding of transcription enhancer factor-1 to the double-stranded polypurine-polypyrimidine element while flanking nucleotides account for interaction of Pur alpha and Pur beta with the purine-rich strand and MSY1 with the complementary pyrimidine-rich strand. Mutations that selectively impaired high affinity single-stranded DNA binding by fibroblast or smooth muscle cell-derived Pur alpha, Pur beta, and MSY1 in vitro, released the cryptic MCAT enhancer from repression in transfected cells. Additional experiments indicated that Pur alpha, Pur beta, and MSY1 also interact specifically, albeit weakly, with double-stranded DNA and with transcription enhancer factor-1. These results are consistent with two plausible models of cryptic MCAT enhancer regulation by Pur alpha, Pur beta, and MSY1 involving either competitive single-stranded DNA binding or masking of MCAT-bound transcription enhancer factor-1.
MeSH term(s)	Amino Acid Motifs ; Animals ; Base Sequence ; Biotinylation ; Cell Line ; Cells, Cultured ; Cyclic AMP Response Element-Binding Protein/metabolism ; DNA/metabolism ; DNA, Single-Stranded/metabolism ; DNA-Binding Proteins/metabolism ; Enhancer Elements, Genetic ; Epitopes ; Fibroblasts/metabolism ; Genes, Reporter ; Mice ; Models, Biological ; Molecular Sequence Data ; Muscle, Smooth/cytology ; Muscle, Smooth/metabolism ; Mutation ; Nerve Tissue Proteins ; Nuclear Proteins ; Precipitin Tests ; Protein Binding ; Rabbits ; Rats ; Transcription Factors/metabolism
Chemical Substances	Cyclic AMP Response Element-Binding Protein ; DNA, Single-Stranded ; DNA-Binding Proteins ; Epitopes ; Nerve Tissue Proteins ; Nsep1 protein, mouse ; Nuclear Proteins ; PURB protein, human ; Pura protein, mouse ; Purb protein, mouse ; TEAD1 protein, human ; Transcription Factors ; DNA (9007-49-2)
Language	English
Publishing date	2001-12-21
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, P.H.S.
ZDB-ID	2997-x
ISSN	1083-351X ; 0021-9258
ISSN (online)	1083-351X
ISSN	0021-9258
DOI	10.1074/jbc.M109754200
Database	MEDical Literature Analysis and Retrieval System OnLINE

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