Cloning of the rat Fibroblast growth factor-2 promoter region and its response to mitogenic stimuli in glioma C6 cells
Identifieur interne : 003C70 ( Main/Exploration ); précédent : 003C69; suivant : 003C71Cloning of the rat Fibroblast growth factor-2 promoter region and its response to mitogenic stimuli in glioma C6 cells
Auteurs : K. B. S. Pasumarthi [Canada] ; Y. Jin [Canada] ; P. A. Cattini [Canada]Source :
- Journal of neurochemistry [ 0022-3042 ] ; 1997.
Descripteurs français
- Pascal (Inist)
English descriptors
- KwdEn :
Abstract
Basic fibroblast growth factor (FGF-2) is abundant in the developing and adult brain and has been linked with the origin and growth of neuronal and glial cells. Glial cells produce high levels of FGF-2, stimulating autocrine growth as well as the survival and function of neurons in a paracrine manner. Abnormal levels of FGF-2 have been linked with Alzheimer's, Huntington's, and Parkinson's diseases. Recent evidence has suggested that a component of the mitogenic response of glial cells is exerted on FGF-2 at the transcriptional level. To assess transcriptional regulation of this potent growth factor, we cloned a 1.4-kb genomic fragment containing the rat FGF-2 promoter region. DNA blotting results indicated that the rat FGF-2 gene exists as a single copy in the genome. The promoter region contains no TATA box but appears to rely instead on multiple GC-rich start sites (P0, P1, and P2) for transcription initiation in rat brain as well as C6 glioma cells. One of these sites (P0) was located within four nucleotides of the reported 5' end of the rat brain cDNA and constituted part of a consensus Egr-1 binding site (5'-GCGGGGGCG-3'). Transcription from this site could be stimulated in C6 glioma cells in response to phorbol ester treatment. The induction of a "new" site (Pi) with phorbol ester also suggested a mechanism to explain the discrepancy between the reported "starts" for the ovarian versus brain cDNAs. A hybrid luciferase gene directed by rat FGF-2 5'-flanking DNA (-1,058/+54) was expressed in rat glioma C6, heart myoblast H9c2, and human astrocytoma U87-MG cells after gene transfer. The level of transfected FGF-2 promoter activity was higher in glial cells (C6 and U87-MG) compared with nonglial (H9c2) cells. Also, expression of this hybrid FGF-2/luciferase gene was increased in response to phorbol ester or serum treatment of C6 cells. Deletion analysis revealed the presence of both positive and negative regulatory regions that are involved in the transcriptional control of rat FGF-2 gene by mitogenic stimuli.
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Jin</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Department of Physiology, University of Manitoba</s1><s2>Winnipeg, Manitoba</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ></inist:fA14><country>Canada</country><placeName><settlement type="city">Winnipeg</settlement><region type="state">Manitoba</region></placeName><orgName type="university">Université du Manitoba</orgName></affiliation></author><author><name sortKey="Cattini, P A" sort="Cattini, P A" uniqKey="Cattini P" first="P. A." last="Cattini">P. A. Cattini</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Department of Physiology, University of Manitoba</s1><s2>Winnipeg, Manitoba</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ></inist:fA14><country>Canada</country><placeName><settlement type="city">Winnipeg</settlement><region type="state">Manitoba</region></placeName><orgName type="university">Université du Manitoba</orgName></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">INIST</idno><idno type="inist">97-0192338</idno><date when="1997">1997</date><idno type="stanalyst">PASCAL 97-0192338 INIST</idno><idno type="RBID">Pascal:97-0192338</idno><idno type="wicri:Area/PascalFrancis/Corpus">000F01</idno><idno type="wicri:Area/PascalFrancis/Curation">001022</idno><idno type="wicri:Area/PascalFrancis/Checkpoint">000D97</idno><idno type="wicri:explorRef" wicri:stream="PascalFrancis" wicri:step="Checkpoint">000D97</idno><idno type="wicri:doubleKey">0022-3042:1997:Pasumarthi K:cloning:of:the</idno><idno type="wicri:Area/Main/Merge">004332</idno><idno type="wicri:Area/Main/Curation">003C70</idno><idno type="wicri:Area/Main/Exploration">003C70</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">Cloning of the rat Fibroblast growth factor-2 promoter region and its response to mitogenic stimuli in glioma C6 cells</title><author><name sortKey="Pasumarthi, K B S" sort="Pasumarthi, K B S" uniqKey="Pasumarthi K" first="K. B. S." last="Pasumarthi">K. B. S. Pasumarthi</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Department of Physiology, University of Manitoba</s1><s2>Winnipeg, Manitoba</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ></inist:fA14><country>Canada</country><placeName><settlement type="city">Winnipeg</settlement><region type="state">Manitoba</region></placeName><orgName type="university">Université du Manitoba</orgName></affiliation></author><author><name sortKey="Jin, Y" sort="Jin, Y" uniqKey="Jin Y" first="Y." last="Jin">Y. Jin</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Department of Physiology, University of Manitoba</s1><s2>Winnipeg, Manitoba</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ></inist:fA14><country>Canada</country><placeName><settlement type="city">Winnipeg</settlement><region type="state">Manitoba</region></placeName><orgName type="university">Université du Manitoba</orgName></affiliation></author><author><name sortKey="Cattini, P A" sort="Cattini, P A" uniqKey="Cattini P" first="P. A." last="Cattini">P. A. Cattini</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Department of Physiology, University of Manitoba</s1><s2>Winnipeg, Manitoba</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ></inist:fA14><country>Canada</country><placeName><settlement type="city">Winnipeg</settlement><region type="state">Manitoba</region></placeName><orgName type="university">Université du Manitoba</orgName></affiliation></author></analytic><series><title level="j" type="main">Journal of neurochemistry</title><title level="j" type="abbreviated">J. neurochem.</title><idno type="ISSN">0022-3042</idno><imprint><date when="1997">1997</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Journal of neurochemistry</title><title level="j" type="abbreviated">J. neurochem.</title><idno type="ISSN">0022-3042</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Cell line</term><term>Fibroblast growth factor</term><term>Glial cell</term><term>In vitro</term><term>Molecular cloning</term><term>Promoter</term><term>Rat</term><term>Regulation(control)</term><term>Regulatory sequence</term><term>Transcription</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Facteur croissance fibroblaste</term><term>Cellule gliale</term><term>In vitro</term><term>Lignée cellulaire</term><term>Clonage moléculaire</term><term>Promoteur</term><term>Rat</term><term>Transcription</term><term>Régulation</term><term>Séquence régulatrice</term><term>Lignée C6</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Basic fibroblast growth factor (FGF-2) is abundant in the developing and adult brain and has been linked with the origin and growth of neuronal and glial cells. Glial cells produce high levels of FGF-2, stimulating autocrine growth as well as the survival and function of neurons in a paracrine manner. Abnormal levels of FGF-2 have been linked with Alzheimer's, Huntington's, and Parkinson's diseases. Recent evidence has suggested that a component of the mitogenic response of glial cells is exerted on FGF-2 at the transcriptional level. To assess transcriptional regulation of this potent growth factor, we cloned a 1.4-kb genomic fragment containing the rat FGF-2 promoter region. DNA blotting results indicated that the rat FGF-2 gene exists as a single copy in the genome. The promoter region contains no TATA box but appears to rely instead on multiple GC-rich start sites (P<sub>0</sub>, P<sub>1</sub>, and P<sub>2</sub>) for transcription initiation in rat brain as well as C6 glioma cells. One of these sites (P<sub>0</sub>) was located within four nucleotides of the reported 5' end of the rat brain cDNA and constituted part of a consensus Egr-1 binding site (5'-GCGGGGGCG-3'). Transcription from this site could be stimulated in C6 glioma cells in response to phorbol ester treatment. The induction of a "new" site (P<sub>i</sub>) with phorbol ester also suggested a mechanism to explain the discrepancy between the reported "starts" for the ovarian versus brain cDNAs. A hybrid luciferase gene directed by rat FGF-2 5'-flanking DNA (-1,058/+54) was expressed in rat glioma C6, heart myoblast H9c2, and human astrocytoma U87-MG cells after gene transfer. The level of transfected FGF-2 promoter activity was higher in glial cells (C6 and U87-MG) compared with nonglial (H9c2) cells. Also, expression of this hybrid FGF-2/luciferase gene was increased in response to phorbol ester or serum treatment of C6 cells. Deletion analysis revealed the presence of both positive and negative regulatory regions that are involved in the transcriptional control of rat FGF-2 gene by mitogenic stimuli.</div></front></TEI><affiliations><list><country><li>Canada</li></country><region><li>Manitoba</li></region><settlement><li>Winnipeg</li></settlement><orgName><li>Université du Manitoba</li></orgName></list><tree><country name="Canada"><region name="Manitoba"><name sortKey="Pasumarthi, K B S" sort="Pasumarthi, K B S" uniqKey="Pasumarthi K" first="K. B. S." last="Pasumarthi">K. B. S. Pasumarthi</name></region><name sortKey="Cattini, P A" sort="Cattini, P A" uniqKey="Cattini P" first="P. A." last="Cattini">P. A. Cattini</name><name sortKey="Jin, Y" sort="Jin, Y" uniqKey="Jin Y" first="Y." last="Jin">Y. Jin</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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