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

Zhang, LM Eggers-Schumacher, G Schoffl, F Prandl, R

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

24

INGLESE

Article

Plant Sciences","Animal & Plant Sciences

PLANT JOURNAL

0960-7412 → ACNP

28

2

2001

217 - 223

ISI

0960-7412(200110)28:2<217:AOHTFB>2.0.ZU;2-U

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.