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Identification of a transcriptional initiator element in the cytochrome c oxidase subunit Vb promoter which binds to transcription factors NF-E1 (YY-1, delta) and Sp1
We have mapped the basal promoter activity of the mouse cytochrome c oxidase (COX) subunit Vb gene to the -17 to +20 region which contains two putative ets binding sites flanking an NF-E1 site fused to an Sp1 site. A 17-nucleotide sequence flanking the major transcription start site (-8 to +9), refe...
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Published in: | The Journal of biological chemistry 1993-02, Vol.268 (6), p.4188-4196 |
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Main Authors: | , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
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Summary: | We have mapped the basal promoter activity of the mouse cytochrome c oxidase (COX) subunit Vb gene to the -17 to +20 region
which contains two putative ets binding sites flanking an NF-E1 site fused to an Sp1 site. A 17-nucleotide sequence flanking
the major transcription start site (-8 to +9), referred to as 17Inr (initiator sequence) was able to drive CAT activity in
3T3 cells to a level comparable to the construct containing sequences -17 to +20. This suggests that the 17Inr sequence contains
the initiator activity. The 17Inr contains a pyrimidine-rich sequence, commencing with a CA that corresponds to the major
transcription start site. Primer extension of RNA from transfected cells demonstrated that transcription initiation with the
17Inr template occurs at a site identical to the endogenous gene. DNA-protein binding by gel mobility shift and methylation
interference analyses indicated that the pyrimidine-rich sequence immediately flanking the transcription start site consists
of an NF-E1 factor binding motif with an overlapping upstream Sp1 binding site. A 13-nucleotide sequence, 13Inr (-4 to +9),
which retains the NF-E1 binding activity but does not bind Sp1, was able to promote chloramphenicol acetyltransferase gene
expression at levels similar to the 17Inr sequence, suggesting that NF-E1 factor binding is critical for initiator function.
Finally, using an in vitro transcription system from Drosophila embryos we demonstrate that NF-E1 is necessary for transcription
activation of both the 17Inr and the 13Inr initiator templates. Thus NF-E1 binding appears to be important for basal promoter
function of the mouse COXVb gene. |
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ISSN: | 0021-9258 1083-351X |
DOI: | 10.1016/S0021-9258(18)53596-1 |