DeLisle A J, Ferl R J
Department of Vegetable Crops, University of Florida, Gainesville 32611.
Plant Cell. 1990 Jun;2(6):547-57. doi: 10.1105/tpc.2.6.547.
Protein-DNA interaction at an inverted repeat of the sequence 5'-GTGG-3' (G-box) has been associated with the transcription of several plant genes [Giuliano, G., et al. (1988). Proc. Natl. Acad. Sci. USA 85, 7089-7093; Ferl, R.J., and Laughner, B.H. (1989). Plant Mol. Biol. 12, 357-366; Schulze-Lefert, P., et al. (1989). EMBO J. 8, 651-656]. We characterized the binding of the Arabidopsis G-box binding factor (GBF) from whole-cell extracts and fractionated extracts to the G-box of alcohol dehydrogenase (Adh) using gel mobility shift assays. DNase I footprinting localized the region of GBF/G-box interaction to two sites, one apparent high-affinity binding site (-227 to -201) and a possible low-affinity binding site (-193 to -182). DNA-protein cross-linking demonstrated that the G-box is bound by proteins of two sizes, 31 kilodaltons and 18 kilodaltons. In addition, we found that in vitro the interaction of GBF from Arabidopsis suspension cultures or leaves with the Adh G-box is indistinguishable, and that there is evidence of multiple protein-protein interactions.
序列5'-GTGG-3'（G盒）的反向重复序列处的蛋白质-DNA相互作用与几种植物基因的转录相关[朱利亚诺，G.等人（1988年）。美国国家科学院院刊85，7089-7093；费尔，R.J.和劳夫纳，B.H.（1989年）。植物分子生物学12，357-366；舒尔茨-勒费特，P.等人（1989年）。欧洲分子生物学组织杂志8，651-656]。我们使用凝胶迁移率变动分析，从全细胞提取物和分级提取物中鉴定了拟南芥G盒结合因子（GBF）与乙醇脱氢酶（Adh）的G盒的结合。DNase I足迹分析将GBF/G盒相互作用区域定位到两个位点，一个明显的高亲和力结合位点（-227至-201）和一个可能的低亲和力结合位点（-193至-182）。DNA-蛋白质交联表明，G盒与两种大小的蛋白质结合，31千道尔顿和18千道尔顿。此外，我们发现，在体外，来自拟南芥悬浮培养物或叶片的GBF与Adh G盒的相互作用没有区别，并且有多种蛋白质-蛋白质相互作用的证据。
