Novel alternative promoters of mouse glial cell line-derived neurotrophic factor gene.
We previously isolated cDNA and genomic DNA of the mouse glial cell line-derived neurotrophic factor (GDNF) gene and found that the gene consists of three exons. Recently, it was suggested that an alternative promoter exists within intron 1 of the human GDNF gene, but this has not been confirmed. Novel cDNA clones of the mouse GDNF gene were isolated by 5'-rapid amplification of cDNA ends from postnatal day-14 striatum. A novel exon, containing 351 nucleotides, exists between exon 1 and exon 3 (referred to as exon 2 in our previous report). Luciferase reporter assay showed that a core promoter for the novel exon 2 requires its 5'-untranslated region. Primer extension analysis and reverse transcription-PCR identified another novel transcript that starts 39 bp upstream of exon 3, and the core promoter activity exists within a region containing putative Sp1 sites. Although the core promoters for the novel exons are different from those previously identified, transcripts derived from each promoter coincidentally increased with interleukin-1beta or tumor necrosis factor-alpha stimulation. Gel retardation assays suggested that the NF-kappaB binding site in intron 1 would be involved in the cytokine response of the mouse GDNF gene.
Tanaka
M
M
Laboratory for Genes of Motor Systems, Bio-Mimetic Control Research Program, The Institute of Physical and Chemical Research Center (RIKEN), Moriyama, 463-0003, Nagoya, Japan.
Ito
S
S
Kiuchi
K
K
D000818:Animals; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism*; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000378:metabolism; D005091:Exons / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects; D051100:Glial Cell Line-Derived Neurotrophic Factor; D007375:Interleukin-1 / Q000494:pharmacology; D007438:Introns / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism; D017072:Neostriatum / Q000166:cytology / Q000378:metabolism; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins / Q000235:genetics*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects / Q000235:genetics*; D014407:Tumor Cells, Cultured
C495040:Gdnf protein, mouse; D051100:Glial Cell Line-Derived Neurotrophic Factor; D007375:Interleukin-1; D016328:NF-kappa B; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D004247:DNA