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Brain Res Mol Brain Res. 1999 Jun 8;69(2):209-22.

Characterization of a promoter for the human glial cell line-derived neurotrophic factor gene.

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  • 1Department of Molecular and Cellular Biochemistry, Center for Biological Research, Neurobiology Unit, Roche Bioscience, M/S R2-101, 3401 Hillview Ave., Palo Alto, CA, 94304-1397, USA. preston.baeker@roche.com

Abstract

To address the regulation of glial cell line-derived neurotrophic factor (GDNF) gene expression, we have isolated 5' extended cDNAs, cloned the human GDNF gene, and characterized the promoter. GDNF-encoding 5' extended cDNAs containing a novel exon were isolated via reverse transcription-polymerase chain reaction (RT-PCR) of mRNA from human fetal kidney and adult skeletal muscle. The GDNF gene was isolated from a human genomic library in a P1 bacteriophage vector. Analysis of the 5' flanking sequence revealed a promoter that lacks a CCAAT-box motif and is GC rich. Consensus binding sites for a variety of transcription factors have been identified in the promoter. Promoter/reporter plasmids have been constructed by fusion of the promoter and a portion of exon I to a luciferase gene. The promoter/reporter construct and a number of promoter deletions were transiently transfected into two human cell lines known to express GDNF. Multiple enhancer and silencer regions were revealed as well as a minimal promoter sufficient for basal transcription. Finally, a RT-PCR assay, specific for transcripts originating from this GDNF promoter, was developed and used to show that this promoter is active in vivo. The results suggest GDNF is regulated in a complex manner.

Copyright 1999 Elsevier Science B.V.

PMID:
10366742
[PubMed - indexed for MEDLINE]
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