Pedersen T J, Arwood L J, Spiker S, Guiltinan M J, Thompson W F
Department of Botany, North Carolina State University, Raleigh 27695-7612.
Plant Mol Biol. 1991 Jan;16(1):95-104. doi: 10.1007/BF00017920.
AT-rich sequences in the 5' flanking regions of several plant genes have been shown to bind nuclear proteins, but the nature of these proteins has remained largely unknown. We report here that certain plant high mobility group (HMG) chromosomal proteins can interact specifically (in the presence of excess non-specific competitor) with AT-rich sequences located upstream of the pea ferredoxin 1 gene (Fed-1) and a member of the wheat Em gene family. Binding was observed with highly purified preparations of HMGa or HMGb, but not with HMGc or HMGd. HMG-DNA complexes were similar to one of the two types of Fed-1 complexes we observed previously using pea nuclear extracts [7]. HMG binding to the Fed-1 DNA was localized to a region containing AT-rich sequences; very similar sequences are present 5' to Em and several other plants genes. Such sequences have been shown to bind unidentified nuclear proteins in a number of these systems. Binding experiments with a synthetic oligo (dA).oligo (dT) probe and competition experiments with synthetic DNA polymers suggest that HMG binding may depend upon structural features of AT-rich DNA rather than being sequence-specific. We discuss the implications of these findings and suggest a role for HMG binding which is consistent with previous evidence linking HMGs with transcriptionally competent chromatin.
几个植物基因5'侧翼区域富含AT的序列已被证明能结合核蛋白,但这些蛋白的性质在很大程度上仍不清楚。我们在此报告,某些植物高迁移率族(HMG)染色体蛋白能(在存在过量非特异性竞争者的情况下)与位于豌豆铁氧还蛋白1基因(Fed-1)上游的富含AT的序列以及小麦Em基因家族的一个成员特异性相互作用。在高度纯化的HMGa或HMGb制剂中观察到了结合,但在HMGc或HMGd中未观察到。HMG-DNA复合物类似于我们先前使用豌豆核提取物观察到的Fed-1复合物的两种类型之一[7]。HMG与Fed-1 DNA的结合定位于一个含有富含AT序列的区域;在Em和其他几个植物基因的5'端存在非常相似的序列。在许多这些系统中,已证明此类序列能结合未鉴定的核蛋白。用合成寡聚体(dA)·寡聚体(dT)探针进行的结合实验以及用合成DNA聚合物进行的竞争实验表明,HMG结合可能取决于富含AT的DNA的结构特征,而不是序列特异性。我们讨论了这些发现的含义,并提出了HMG结合的作用,这与先前将HMG与转录活性染色质联系起来的证据一致。