Early growth response transcription factor EGR-1 regulates Galphaq gene in megakaryocytic cells.

Early growth response transcription factor EGR-1 regulates Galphaq gene in megakaryocytic cells. Research Abstract Details 

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Early growth response transcription factor EGR-1 regulates Galphaq gene in megakaryocytic cells. Abstract Text:

G Jalagadugula,D N Dhanasekaran,S Kim,S P Kunapuli,A K Rao,

BACKGROUND: Galphaq (Gene GNAQ) plays a major role in platelet signal transduction but little is known regarding its transcriptional regulation. OBJECTIVES: We studied Galphaq promoter activity using luciferase reporter gene assays in human erythroleukemia (HEL) cells treated with phorbol 12-myristate 13-acetate (PMA) for 24 h to induce megakaryocytic transformation. METHODS AND RESULTS: PMA-treated HEL cells showed enhanced Galphaq expression. Reporter (luciferase) gene studies on 5' upstream construct (up to -116 bp from ATG) revealed a negative regulatory site at -238/-202 and two positive sites at -203/-138 and -1116/-731. The positive regulatory region -203/-138 contained overlapping Sp1/AP-2/EGR-1 consensus sites. Gel shift studies on Galphaq oligonucleotides 1 (-203/-175) and 2 (-174/-152) using HEL cell extracts demonstrated protein binding that was due to early growth response factor EGR-1 at two sites. Mutations in either EGR-1 site markedly decreased the gene activity, indicating functional relevance. Mutation of consensus E-Box motif (-185/-180) had no effect. Reduction in the expression of endogenous EGR-1 with antisense oligonucleotide to EGR-1 inhibited PMA-induced Galphaq transcription. Correspondingly, Egr-1 deficient mouse platelets also showed approximately 50% reduction in the Galphaq expression relative to wild-type platelets. CONCLUSIONS: These studies suggest that Galphaq gene is regulated during PMA-induced megakaryocytic differentiation by EGR-1, an early growth response transcription factor that regulates a wide array of genes and plays a major role in diverse activities, including cell proliferation, differentiation and apoptosis, and in vascular response to injury and atherosclerosis.

Early growth response transcription factor EGR-1 regulates Galphaq gene in megakaryocytic cells. Publishing Authors By Initials

G Jalagadugula,DN Dhanasekaran,S Kim,SP Kunapuli,AK Rao,

For similar investigative techniques: genetic techniques: gene transfer techniques: transfection research abstracts see: investigative techniques: genetic techniques: gene transfer techniques: transfection research

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Early growth response transcription factor EGR-1 regulates Galphaq gene in megakaryocytic cells. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Journal of thrombosis and haemostasis : JTH

VOLUME: 4

Page Numbers: 2678-86

Journal Abbreviation: J. Thromb. Haemost.

ISSN: 1538-7933

DAY: 22

MONTH: 09

YEAR: 2006

Early growth response transcription factor EGR-1 regulates Galphaq gene in megakaryocytic cells. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 101170508

Early growth response transcription factor EGR-1 regulates Galphaq gene in megakaryocytic cells. Keywords Mesh Terms:

KEYWORDS: Transfection

MESH TERMS: pharmacology

Chemical & Substance for Abstract: Early growth response transcription factor EGR-1 regulates Galphaq gene in megakaryocytic cells. Information

Substance Name: GTP-Binding Protein alpha Subunits, Gq-G

Registry Number: EC 3.6.1.46

Grant and Affiliation Information for Early growth response transcription factor EGR-1 regulates Galphaq gene in megakaryocytic cells.

AFFILIATION: Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA 19140, USA.

Country: England

AGENCY: United States NHLBI

GRANT: R01 HL56724

ACRONYM: HL

MEDLINETA: J Thromb Haemost

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