Cloning and structural organization of the gene encoding the mouse glial cell line-derived neurotrophic factor, GDNF.
cDNA for glial cell line-derived neurotrophic factor (GDNF) was cloned from mouse neonatal brain by the method of 5'-rapid amplification of cDNA end (5'-RACE), and the sequence of it's 5'-untranslated region (5'-UTR) was determined. The mouse GDNF gene was then isolated from a genomic library and analyzed for its nucleotide sequence. In vitro translation analysis indicated that the second ATG codon in an open reading frame is the translation start point. Structural analysis of the isolated clones showed that the GDNF gene was separated into three exons and the actual translation start point was present in the second exon. RNA blot hybridization analysis indicated that the GDNF mRNA is approximately 4.5 kb long. The transcriptional start site in the GDNF gene was determined and a typical TATA box sequence was found in the promoter region. On the other hand, the gene expression of GDNF in C6 glioma cells was transiently induced by treatment with phorbol myristate acetate (PMA), but not by forskolin.
Matsushita
N
N
Laboratory of Genes of Motor Systems, Bio-Mimetic Control Research Program, The Institute of Physical and Chemical Research Center (RIKEN), Moriyama, Nagoya, Japan.
Fujita
Y
Y
Tanaka
M
M
Nagatsu
T
T
Kiuchi
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001923:Brain Chemistry; D003001:Cloning, Molecular; D005576:Colforsin / Q000494:pharmacology; D005091:Exons / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects; D015723:Gene Library; D051100:Glial Cell Line-Derived Neurotrophic Factor; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000235:genetics*; D011233:Precipitin Tests; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D016385:TATA Box / Q000235:genetics; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic / Q000235:genetics; D014407:Tumor Cells, Cultured
C495040:Gdnf protein, mouse; D051100:Glial Cell Line-Derived Neurotrophic Factor; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D005576:Colforsin; D013755:Tetradecanoylphorbol Acetate