Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
4
pubmed:dateCreated
1999-7-15
pubmed:abstractText
Our previous studies have shown that transcription of brain creatine kinase (CKB) mRNA in U87-MG glioblastoma cells is stimulated by a forskolin-mediated increase in cyclic AMP (cAMP) via a pathway involving protein kinase A (PKA) and the activation of Galphas proteins. In this report, we have employed transient transfection to investigate the rat CKB gene elements essential for the cAMP-mediated induction of rat CKB transcription in human U87 cells and have mapped the transcription start site of the induced CKB transcripts. We found that the level of induced transcription from the transfected genomic rat CKB gene was the same whether transcription was driven by 2.9 kb of CKB promoter plus 5' flanking sequence or the 0.2 kb CKB promoter, suggesting that the proximal CKB promoter was essential. Also, the level of induced transcription of the chloramphenicol acetyl transferase (CAT) reporter gene driven by the 2.9 kb CKB promoter was the same as with the 0.2 kb CKB promoter. Analyses of a series of 5' deletions of the 0.2 kb proximal CKB promoter showed that the sequences between -80 bp and +1 bp were essential for the cAMP-mediated induction of CKB transcription, despite the absence of a consensus cAMP response element (CRE) sequence in that region. In agreement, gel mobility shift assays showed that nuclear extracts from U87 cells contained a protein(s) which bound specifically to a [32P]CKB DNA probe containing the -60 bp to +1 bp sequence. Mapping the 5' end of the CKB transcripts showed that the initiation of the cAMP-induced transcription occurred almost exclusively from the downstream transcription start site, apparently under the initiation direction of the nonconsensus (-28) TTAA element and not the consensus (-60) TATAAATA element. The results are discussed with regard to nuclear protein factors which may be involved, and the possible cAMP-mediated increase in CKB transcription during myelinogenesis, since the differentiation of oligodendrocytes has previously been shown to be accelerated by increased intracellular cAMP.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
May
pubmed:issn
0360-4012
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
56
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
371-85
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:10340745-1-Methyl-3-isobutylxanthine, pubmed-meshheading:10340745-Animals, pubmed-meshheading:10340745-Astrocytes, pubmed-meshheading:10340745-Base Sequence, pubmed-meshheading:10340745-Brain, pubmed-meshheading:10340745-Consensus Sequence, pubmed-meshheading:10340745-Creatine Kinase, pubmed-meshheading:10340745-Cyclic AMP, pubmed-meshheading:10340745-DNA Probes, pubmed-meshheading:10340745-Forskolin, pubmed-meshheading:10340745-Gene Expression Regulation, Enzymologic, pubmed-meshheading:10340745-Glioblastoma, pubmed-meshheading:10340745-Humans, pubmed-meshheading:10340745-Introns, pubmed-meshheading:10340745-Nuclear Proteins, pubmed-meshheading:10340745-Promoter Regions, Genetic, pubmed-meshheading:10340745-RNA, Messenger, pubmed-meshheading:10340745-Rats, pubmed-meshheading:10340745-Response Elements, pubmed-meshheading:10340745-TATA Box, pubmed-meshheading:10340745-Transcriptional Activation, pubmed-meshheading:10340745-Transfection, pubmed-meshheading:10340745-Tumor Cells, Cultured
pubmed:year
1999
pubmed:articleTitle
Proximal promoter of the rat brain creatine kinase gene lacks a consensus CRE element but is essential for the cAMP-mediated increased transcription in glioblastoma cells.
pubmed:affiliation
Department of Biology, University of Delaware, Newark 19716, USA.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't