Bound transcription factor suppresses photoproduct formation in the NF-κB promoter

Rita Ghosh, Lakshmi Paniker, David L. Mitchell

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The relationship between purified transcription factor p50 binding and ultraviolet light-induced DNA damage formation in the NF-κB promoter element was investigated. The effect of bound transcription factor on cyclobutane dimer formation was quantified using Maxam-Gilbert analysis of irradiated substrate digested with T4 phage endonuclease V. Two methods were employed for cleaving (6-4) photoproducts. Sites of (6-4) photoproducts cleaved by piperidine showed a general suppression in the presence of bound p50 protein similar to that observed for cyclobutane dimers. In contrast to piperidine, digestion with ultraviolet damage endonuclease (UVDE) from Saccharomyces pombe subsequent to cyclobutane dimer reversal by photolyase displayed a broader spectrum of damaged sites. Whereas some of these sites were suppressed by bound p50 protein, some remained unaffected and one site showed increased (6-4) photoproduct induction. These data illustrate the advantage of UVDE over piperidine for studying (6-4) photoproducts at the sequence level and suggest that this approach may be useful for footprinting transcription factor binding in other promoters.

Original languageEnglish (US)
Pages (from-to)1-5
Number of pages5
JournalPhotochemistry and Photobiology
Volume73
Issue number1
DOIs
StatePublished - Jan 1 2001
Externally publishedYes

Fingerprint

Cyclobutanes
cyclobutane
piperidine
Dimers
Transcription Factors
Endonucleases
dimers
damage
Deoxyribodipyrimidine Photo-Lyase
proteins
saccharomyces
Saccharomyces
Ultraviolet Rays
ultraviolet radiation
DNA Damage
Digestion
induction
Proteins
deoxyribonucleic acid
retarding

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biophysics

Cite this

Bound transcription factor suppresses photoproduct formation in the NF-κB promoter. / Ghosh, Rita; Paniker, Lakshmi; Mitchell, David L.

In: Photochemistry and Photobiology, Vol. 73, No. 1, 01.01.2001, p. 1-5.

Research output: Contribution to journalArticle

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