Linked Life Data

7577965

Source:http://linkedlifedata.com/resource/pubmed/id/7577965

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
42
pubmed:dateCreated
1995-12-14
pubmed:databankReference
pubmed:abstractText
The rat P-450c27/25 (CYP27) gene is expressed as two distinctly sized mRNAs of 2 and 2.3 kb (kilobase). The 2 kb mRNA is the predominant form in the liver with negligible 2.3 kb species. Rat kidney and hepatoma, on the other hand, contain significant levels of the 2.3 kb species. Rat CYP27 gene contains 11 exons of 80-415 nucleotides that are separated by 10 introns of 83 bases to approximately 10 kb. S1 nuclease protection and primer extension analyses using liver RNA showed a prominent 5' terminus 86 nucleotides downstream from the start of exon 2. This site, designated as +1, is the start site for the 2 kb mRNA. 5' RACE analysis of rat kidney and hepatoma RNAs showed the presence of a 5' extended mRNA with a sequence complementary to the Spi2 mRNA. A cryptic TATA box (TTTAAA) is located 24 nucleotides upstream of the 2 kb mRNA transcription initiation site at +1. A 106 bp DNA fragment (sequence -83 to +23) that houses the putative TATA motif forms three differently migrating complexes with nuclear extract from the murine 3T3 cells. DNAse I footprinting and competition with synthetic DNA showed that complex A represents the bound Sp1 factor and complexes B and C are due to unknown factors binding to the -83 to -71 and -20 to -12 sequences, respectively. In vivo transcription analysis using -840/+23 DNA and its 5' deletions cloned in a CAT reporter plasmid suggests that the basal promoter elements are located within sequence -45 to +23 of the gene. Finally, in vitro transcription analysis in HeLa cell nuclear extract showed that intact TTTAAA motif and complex C-forming sequence from this region are essential for transcription initiation at the +1 position of the promoter. Our results demonstrate that the 2 kb mRNA is transcribed as an independent transcript driven by an immediate upstream promoter located within exon 2.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
0006-2960
pubmed:author
pubmed:issnType
Print
pubmed:day
24
pubmed:volume
34
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
13729-42
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:7577965-Animals, pubmed-meshheading:7577965-Base Sequence, pubmed-meshheading:7577965-Blotting, Northern, pubmed-meshheading:7577965-Carcinoma, Hepatocellular, pubmed-meshheading:7577965-Cell Line, pubmed-meshheading:7577965-Cytochrome P-450 Enzyme System, pubmed-meshheading:7577965-DNA Primers, pubmed-meshheading:7577965-DNA-Binding Proteins, pubmed-meshheading:7577965-Deoxyribonuclease I, pubmed-meshheading:7577965-Exons, pubmed-meshheading:7577965-Genes, Reporter, pubmed-meshheading:7577965-Kidney, pubmed-meshheading:7577965-Molecular Sequence Data, pubmed-meshheading:7577965-Promoter Regions, Genetic, pubmed-meshheading:7577965-RNA, Messenger, pubmed-meshheading:7577965-Rats, pubmed-meshheading:7577965-Regulatory Sequences, Nucleic Acid, pubmed-meshheading:7577965-Restriction Mapping, pubmed-meshheading:7577965-Single-Strand Specific DNA and RNA Endonucleases, pubmed-meshheading:7577965-TATA Box, pubmed-meshheading:7577965-Transcription, Genetic, pubmed-meshheading:7577965-Tumor Cells, Cultured
pubmed:year
1995
pubmed:articleTitle
Localization of a transcription promoter within the second exon of the cytochrome P-450c27/25 gene for the expression of the major species of two-kilobase mRNA.
pubmed:affiliation
Department of Animal Biology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia 19104-6046, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.