Identification of the transcription termination site of the mouse nkx-1.2 gene : involvement of sequence-specific factors

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Title
Identification of the transcription termination site of the mouse nkx-1.2 gene : involvement of sequence-specific factors
Author(s)
Seung-Beom Hong; Sun Jung Kim; Moon Jong Noh; Young Mi Lee; Yongsok Kim; Ook Joon Yoo
Bibliographic Citation
Gene, vol. 198, no. 1, pp. 373-378
Publication Year
1997
Abstract
We have identified a transcription termination site in the 3′ flanking region of the mouse nkx-1.2 gene. A downstream transcription regulatory element in the mouse nkx-1.2 gene was characterized by transferring its 3′-fragment into a chloramphenicol acetyl transferase (CAT) expression vector. Analysis of recombinant plasmids transfected into mouse NIH3T3 cells by CAT assay showed the possible region of regulation. There were two direct repeat structures containing poly(dG-dT) · poly(dC-dA) sequences (GT repeats) in this region. The precise location of transcription termination was mapped by nuclease S1 analysis of the transcripts from recombinant plasmids transfected into COSM6 cells. It was approximately 20 nucleotides upstream of the first GT repeat within the 5′ sequences of the first element of the two direct repeats. Gel mobility shift assay and footprinting analysis demonstrated that nuclear DNA binding proteins bound specifically to the sequences where the termination occurred as well as the other sequences in the second element of the direct repeats. Southwestern analysis showed that 90-, 54-, 36- and 15-kDa nuclear proteins bound to the region of the termination. It is possible that one or more of those proteins are involved in blocking the elongation of the mouse nkx-1.2 gene transcript and then result in termination.
Keyword
GT repeatTranscription regulationRNA polymerase II
ISSN
0378-1119
Publisher
Elsevier
DOI
http://dx.doi.org/10.1016/s0378-1119(97)00340-5
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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