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Analysis on the promoter region of exon IV brain‐derived neurotrophic factor in NG108‐15 cells
Author(s) -
Takeuchi Yusuke,
Miyamoto Eishichi,
Fukunaga Kohji
Publication year - 2002
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.2002.01096.x
Subject(s) - exon , microbiology and biotechnology , brain derived neurotrophic factor , promoter , transcription factor , sp1 transcription factor , neurotrophic factors , chemistry , transcription (linguistics) , protein kinase a , transfection , gene isoform , biology , kinase , gene expression , biochemistry , gene , receptor , linguistics , philosophy
We have reported that the nuclear isoforms of Ca 2+ /calmodulin‐dependent protein kinase II (CaM KII) are involved in the expression of the exon IV‐containing brain‐derived neurotrophic factor (BDNF) mRNA. We document here the cis ‐elements and transcription factors responsive to CaM KII inthe activation of the promoter upstream of the exon IV (exon IV‐BDNF promoter). Effects of constitutively active mutants of CaM KIV, MAPK kinase kinase (MEKK) and protein kinase A (PKA) on the exon IV‐BDNF promoter activity were also evaluated by transfection and luciferase assay. The exon IV‐BDNF promoter activity was increased by transfection with CaM KII, MEKK and PKA, but not by CaM KIV. Deletion and mutational analysis of the promoter revealed that the region between nucleotides −56 and −27 was responsive to CaM KII, which contained a CCAAT‐box in the region between nucleotides −56 and −43. Expression of C/EBPβ increased the promoter activity and potentiated the effects of CaM KII. The region between nucleotides −79 and −56 was responsive to MEKK, in which both a GA‐rich sequence and a GC‐box were included. Expression of Sp1 increased the promoter activity, which was further enhanced by transfection with MEKK. The region between nucleotides −43 and −27 was responsive to both PKA and CaM KII, but the transcription factors involved in the region remained unclear. These results suggest that the promoter of the exon IV‐BDNF is at least regulated by CaM KII, MEKK and PKA, and that C/EBP/β and Sp1 are potential transcription factors activated by CaM KII and MEKK, respectively.