Restoration of TATA-dependent transcription in a heat-inactivated extract of tobacco nuclei by recombinant TATA-binding protein (TBP) from tobacco

Restoration of TATA-dependent transcription in a heat-inactivated extract of tobacco nuclei by... We isolated a complementary DNA (cDNA) that encoded a TATA-binding protein (TBP) from a cDNA library of tobacco (Nicotiana tabacum) suspension-cultured cells (BY-2). A comparison among deduced amino acid sequences of plant TBPs revealed the presence of a long conserved region within the amino acid sequence of the TBP. Genomic Southern analysis revealed that tobacco TBP (tTBP) is encoded by only a small number of copies of a gene in the tobacco genome. Addition of recombinant tTBP to an extract of tobacco nuclei (TNE) enhanced the basal transcriptional activity in vitro. This result indicates that the level of tTBP is a rate-limiting factor for basal transcriptional activity in TNE. We subsequently succeeded in the functional complementation of TATA-dependent initiation of transcription that was associated with a plant promoter in a homologous plant system. Addition of bacterially expressed recombinant tTBP to a heat-inactivated TNE restored transcriptional activity, as did the addition of human TBP. Moreover, heating of the recombinant tTBP eliminated its ability to restore transcriptional activity. It appears that the heat inactivation of TNE was caused by the heat inactivation of tTBP in TNE. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Plant Molecular Biology Springer Journals

Restoration of TATA-dependent transcription in a heat-inactivated extract of tobacco nuclei by recombinant TATA-binding protein (TBP) from tobacco

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Publisher
Kluwer Academic Publishers
Copyright
Copyright © 1997 by Kluwer Academic Publishers
Subject
Life Sciences; Biochemistry, general; Plant Sciences; Plant Pathology
ISSN
0167-4412
eISSN
1573-5028
D.O.I.
10.1023/A:1005759521285
Publisher site
See Article on Publisher Site

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