Abstract:
:The neprilysin gene is composed of three distinct 5' noncoding exons which can be joined to the first coding exon to generate multiple mRNA species, all encoding the same protein. Genomic fragments containing upstream sequences of each of these three noncoding exons from the rat neprilysin gene were subcloned in the promoterless vector pXp1, which contains the luciferase reporter gene. Expression was compared between a neprilysin positive human spinal cord cell line, HSC-2, and neprilysin negative lines MCF-7, a human breast adenocarcinoma cell line and Hep G2, a human liver carcinoma cell line. The first and second promoter regions showed high activity in the positive cell line, but low activity in the negative cell lines. An analysis of the exon 1 promoter region showed that the proximal 85 nucleotides exhibited basal promoter activity. An enhancer-like sequence was found to be located within a 22-bp fragment located at -136 to -115. Scanning mutagenesis of a 29-bp fragment containing the enhancer-like sequence showed that changes in each 5- or 6-bp segment throughout this fragment decreased activity; however, mutations of the segment encompassing positions 19 to 24 eliminated >98% of the promoter activity. Binding of nuclear proteins from HSC-2 cells to this 29-bp fragment was observed by gel shift analysis. The ability of mutations within the 29-bp fragment to affect enhancer activity correlated with the ability of these mutant oligonucleotides to compete for the wild-type sequence in gel shift assays.
journal_name
Arch Biochem Biophysjournal_title
Archives of biochemistry and biophysicsauthors
Li C,Hersh LBdoi
10.1006/abbi.1998.0855subject
Has Abstractpub_date
1998-10-01 00:00:00pages
189-95issue
1eissn
0003-9861issn
1096-0384pii
S0003-9861(98)90855-4journal_volume
358pub_type
杂志文章abstract::Membrane dielectric as a primary basis for effects of ethanol was examined in synaptic plasma membranes (SPM) of genetically selected ethanol-sensitive long-sleep (LS) and ethanol-resistant short-sleep (SS) mice. Multifrequency phase and modulation of fluorometry of diphenylhexatriene (DPH) was used to resolve structu...
journal_title:Archives of biochemistry and biophysics
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journal_title:Archives of biochemistry and biophysics
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journal_title:Archives of biochemistry and biophysics
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journal_title:Archives of biochemistry and biophysics
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更新日期:2009-04-01 00:00:00
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journal_title:Archives of biochemistry and biophysics
pub_type: 杂志文章
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更新日期:1985-07-01 00:00:00