TY - JOUR
T1 - Analysis of the human GDNF gene reveals an inducible promoter, three exons, a triplet repeat within the 3'-UTR and alternative splice products
AU - Grimm, Lena
AU - Holinski-Feder, Elke
AU - Teodoridis, Jens
AU - Scheffer, Beatrix
AU - Schindelhauer, Dirk
AU - Meitinger, Thomas
AU - Ueffing, Marius
N1 - Funding Information:
The expression vector pBC140 was kindly provided by Dorian Bevec and pAH1409 was a generous gift of Albrecht Sippel. We thank Dr Thomas Gasser for DNA samples of patients with neurodegenerative diseases. We are grateful to Drs Clemens Suter-Crazzolara, Dirk Eick, Ursula Just and Marc Billaud for valuable discussions and advice and for critical reading of the manuscript. We want to thank Georg W. Bornkamm for continuous support. This research was supported by grant 01KW9605/4 from the German BMBF.
PY - 1998/11
Y1 - 1998/11
N2 - Glial cell line-derived neurotrophic factor (GDNF), a distant member of the TGF-β superfamily, is a survival factor for various neurons, making it a potential therapeutic agent for neurodegenerative disorders. Here we present the genomic structure and characterization of the promoter of the human GDNF (hGDNF) gene. It contains three exons coding for a cDNA of 4.6 kb including large 5'- and 3'-untranslated regions (UTRs). The 3'-UTR contains a polymorphic AGG repeat that appears not to be expanded in patients suffering from different neurodegenerative disorders. RT-PCR results in at least three different hGDNF transcripts including one that lacks exon 2. Transient expression experiments reveal that exon 2 is essential for proper cellular processing to yield a secreted form of hGDNF, whereas expression of exon 3 alone is sufficient to code for a mature form of hGDNF retained within the cell. Our data show that the hGDNF gene is driven by a TATA-containing promoter preceding exon 1. A second promoter element has been mapped to a region 5' of exon 2. Both promoters are in close proximity to CpG islands covering exons 1 and 2. Using luciferase as a reporter gene, the TATA-containing hGDNF promoter facilitates a 20- to 40-fold increase in transcription when compared with a corresponding promoterless construct, whereas the second promoter confers only weak activity. Furthermore, fibroblast growth factor 2, tetradecanoyl 12-phorbol acetate, an inflammatory agent, and cAMP increase promoter activity, suggesting that GDNF transcriptional regulation is a target of exogenous signals.
AB - Glial cell line-derived neurotrophic factor (GDNF), a distant member of the TGF-β superfamily, is a survival factor for various neurons, making it a potential therapeutic agent for neurodegenerative disorders. Here we present the genomic structure and characterization of the promoter of the human GDNF (hGDNF) gene. It contains three exons coding for a cDNA of 4.6 kb including large 5'- and 3'-untranslated regions (UTRs). The 3'-UTR contains a polymorphic AGG repeat that appears not to be expanded in patients suffering from different neurodegenerative disorders. RT-PCR results in at least three different hGDNF transcripts including one that lacks exon 2. Transient expression experiments reveal that exon 2 is essential for proper cellular processing to yield a secreted form of hGDNF, whereas expression of exon 3 alone is sufficient to code for a mature form of hGDNF retained within the cell. Our data show that the hGDNF gene is driven by a TATA-containing promoter preceding exon 1. A second promoter element has been mapped to a region 5' of exon 2. Both promoters are in close proximity to CpG islands covering exons 1 and 2. Using luciferase as a reporter gene, the TATA-containing hGDNF promoter facilitates a 20- to 40-fold increase in transcription when compared with a corresponding promoterless construct, whereas the second promoter confers only weak activity. Furthermore, fibroblast growth factor 2, tetradecanoyl 12-phorbol acetate, an inflammatory agent, and cAMP increase promoter activity, suggesting that GDNF transcriptional regulation is a target of exogenous signals.
UR - http://www.scopus.com/inward/record.url?scp=0031794590&partnerID=8YFLogxK
U2 - 10.1093/hmg/7.12.1873
DO - 10.1093/hmg/7.12.1873
M3 - Article
C2 - 9811930
AN - SCOPUS:0031794590
SN - 0964-6906
VL - 7
SP - 1873
EP - 1886
JO - Human Molecular Genetics
JF - Human Molecular Genetics
IS - 12
ER -