AINTEGUMENTA utilizes a mode of DNA recognition distinct from that used by proteins containing a single AP2 domain
AINTEGUMENTA utilizes a mode of DNA recognition distinct from that used by proteins containing a single AP2 domain
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DOI：
10.1093/nar/gkg292
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发表时间：
2003-04-01
影响因子：
14.9
通讯作者：
Krizek, BA
中科院分区：
文献类型：
--
作者：
Krizek, BA
拟南芥蛋白AINTEGUMENTA（ANT）是花发育过程中器官生长的重要调节因子。ANT是植物特异性转录因子AP 2/ERF家族的AP 2亚类的成员。这些蛋白质包含一个或两个称为AP 2结构域的DNA结合结构域拷贝。在这里，它表明，ANT可以作为一个转录激活酵母通过结合到一个共识ANT结合位点。该活性被用作遗传筛选的基础，以鉴定对ANT的DNA结合能力至关重要的氨基酸。鉴定了在ANT结合位点控制下显示报告基因活化减少或无活化的突变体。在筛选中鉴定的突变以及额外的定点突变表明，AP 2亚家族成员的DNA识别模式与ERF蛋白不同。令人惊讶的是，ANT的每个AP 2结构域似乎使用不同的氨基酸来接触DNA。几个接头突变的鉴定表明，该序列在DNA上每个AP 2结构域的定位中起作用，或直接与DNA接触。
The Arabidopsis protein AINTEGUMENTA ( ANT) is an important regulator of organ growth during flower development. ANT is a member of the AP2 subclass of the AP2/ERF family of plant-specific transcription factors. These proteins contain either one or two copies of a DNA-binding domain called the AP2 domain. Here, it is shown that ANT can act as a transcriptional activator in yeast through binding to a consensus ANT-binding site. This activity was used as the basis for a genetic screen to identify amino acids that are critical for the DNA binding ability of ANT. Mutants that showed reduced or no activation of a reporter gene under the control of ANT-binding sites were identified. The mutations identified in the screen as well as additional site-directed mutations suggest that the mode of DNA recognition by members of the AP2 subfamily is distinct from that of ERF proteins. Surprisingly, it appears that each AP2 domain of ANT uses different amino acids to contact DNA. Identification of several linker mutations argues that this sequence acts in the positioning of each AP2 domain on the DNA or makes direct DNA contacts.