Open AccessFunctional identification of the promoter for the gene encoding the alpha subunit of calcium/calmodulin-dependent protein kinase II.
Author(s) -
Norma J. Olson,
Thierry Massé,
Tatsuo Suzuki,
Jing Chen,
Daniel Alam,
Paul Kelly
Publication year - 1995
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.92.5.1659
Subject(s) - microbiology and biotechnology , biology , chloramphenicol acetyltransferase , transcription (linguistics) , tata box , promoter , response element , reporter gene , g alpha subunit , gene , scn3a , protein kinase a , transfection , protein subunit , gene expression , kinase , genetics , linguistics , philosophy
To examine the expression of the alpha subunit of calcium/calmodulin-dependent protein kinase II, various 5' flanking genomic sequences were inserted into a chloramphenicol acetyltransferase (CAT) reporter plasmid and CAT enzyme activities were analyzed in transfected NB2a neuroblastoma cells and mRNA transcription was analyzed by nuclease protection assays. A core promoter was identified which contained an essential TATA element located 162 nt 5' to the transcription start site. Sequences 3' to the transcription start site, as well as 5' to the TATA element, increased levels of CAT activity in transfected cells. The alpha-subunit gene promoter displayed higher CAT activities, relative to a simian virus 40 promoter, in transfected neuronal cell lines than in nonneuronal cell lines. Results also suggested that sequence surrounding the natural alpha-gene transcription initiation site may be important for targeting transcription initiation 162 nt downstream of its TATA element.