Sp1 activation of RNA polymerase II transcription complexes involves a heat-labile DNA-binding component.

P. J. Farnham, M. M. Cornwell

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

We have identified a component of the eukaryotic RNA polymerase II transcriptional machinery that is more heat-labile than TFIID. DHFR transcriptional activity was severely reduced in 40 degrees C heat-treated extracts in which TFIID was fully active. This heat-labile activity was required for the transcription of both TATA box and non-TATA box promoters that are activated by the transcription factor Sp1. Gel mobility shifts indicated that Sp1 DNA binding activity was heat-labile, and the addition of purified Sp1 to 40 degrees C heat-treated extracts fully restored DHFR transcriptional activity. In contrast, the addition of Sp1 to 47 degrees C heat-treated extract did not result in transcriptional activity from the DHFR promoter. We conclude that reduction in Sp1 DNA binding activity is partially responsible for the heat-sensitive loss of DHFR transcriptional activity, but that a second essential activity is also inactivated by 47 degrees C heat-treatment. The discovery of this heat-labile component of Sp1 activation has two important implications in the analysis of transcriptional regulation. First, it demonstrates that heat-treated extracts are not appropriate for examination of the involvement of TFIID in the transcription of Sp1-activated promoters. Second, it explains the previously reported low-temperature optima for transcription from the DHFR promoter and demonstrates that transcriptional studies of Sp1-activated promoters should not be performed at 30 degrees C.

Original languageEnglish (US)
Pages (from-to)137-148
Number of pages12
JournalGene Expression
Volume1
Issue number2
StatePublished - May 1991
Externally publishedYes

Fingerprint

RNA Polymerase II
Hot Temperature
DNA
Transcription Factor TFIID
Sp1 Transcription Factor
TATA Box
Gels
Temperature

ASJC Scopus subject areas

  • Genetics

Cite this

Sp1 activation of RNA polymerase II transcription complexes involves a heat-labile DNA-binding component. / Farnham, P. J.; Cornwell, M. M.

In: Gene Expression, Vol. 1, No. 2, 05.1991, p. 137-148.

Research output: Contribution to journalArticle

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