Analysis of heat-shock transcription factor-DNA binding in Arabidopsis suspension cultures by UV laser crosslinking

Citation
Lm. Zhang et al., Analysis of heat-shock transcription factor-DNA binding in Arabidopsis suspension cultures by UV laser crosslinking, PLANT J, 28(2), 2001, pp. 217-223
Citations number
24
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Plant Sciences","Animal & Plant Sciences
Journal title
PLANT JOURNAL
ISSN journal
0960-7412 → ACNP
Volume
28
Issue
2
Year of publication
2001
Pages
217 - 223
Database
ISI
SICI code
0960-7412(200110)28:2<217:AOHTFB>2.0.ZU;2-U
Abstract
Crosslinking techniques are important in examining protein-DNA interactions in living cells. Formaldehyde and UV light emitted by conventional lamps a re the most commonly used crosslinking agents. The crosslinking step is fol lowed by immunoprecipitation of specific protein-DNA adducts, and by analys is and quantification of the co-immunoprecipitated DNA. There are a few rep orted cases of fruitful in vivo protein-DNA crosslinking experiments, but n ot in plants. In this report, we analyse the binding of heat-shock transcri ption factor (HSF) to heat-shock promoters in intact Arabidopsis cells. Eff icient protein-DNA crosslinking by irradiation of Arabidopsis suspension cu lture tissue was achieved using UV laser pulses. In addition, methods for i mmunoprecipitation and detection of the coimmunoprecipitated DNA are report ed. Our results suggest that Arabidopsis HSFs immunoreactive for HSF1 antib odies bind constitutively to the HSP18.2 gene.