Jeong Mi-Jeong, Shih Ming-Che
Department of Biological Sciences, 204 Chemistry Building, University of Iowa, Iowa City, IA 52242, USA.
Biochem Biophys Res Commun. 2003 Jan 10;300(2):555-62. doi: 10.1016/s0006-291x(02)02892-9.
We have previously identified a cis-acting element, named the XXIII box, that is essential for light-regulated expression of the nuclear gene GAPB, which encodes the B subunit of chloroplast glyceraldehyde-3-phosphate dehydrogenase from Arabidopsis thaliana. Examination of the sequences indicated that there are two GATA motifs within the XXIII box. Based on the degree of the amino-acid sequence identity in the DNA binding domains, we divided the 25 GATA factors encoded in the Arabidopsis genome into three classes. We chose GATA-1 and GATA-20 from Class I and Class II, which include the majority of GATA factors, for overexpression in an Escherichia coli expression system. Gel mobility shift assays showed that GATA-1, but not GATA-20, binds specifically to the two GATA motifs within the XXIII fragment. In addition, we showed that GATA-1 could also bind specifically to a cis-acting element in the promoter of the GAPA gene, which is coordinately regulated by light with the GAPB gene. Based on these results, we propose that light controls the expression of GAPA and GAPB genes in part by regulating the binding of the same transcription factor at their GATA motifs.
我们之前鉴定出了一种顺式作用元件,命名为XXIII框,它对于核基因GAPB的光调控表达至关重要,GAPB编码拟南芥叶绿体甘油醛-3-磷酸脱氢酶的B亚基。对序列的检查表明,XXIII框内有两个GATA基序。根据DNA结合结构域中氨基酸序列的同一性程度,我们将拟南芥基因组中编码的25个GATA因子分为三类。我们从I类和II类中选择了GATA-1和GATA-20进行在大肠杆菌表达系统中的过表达,I类和II类包含了大多数GATA因子。凝胶迁移率变动分析表明,GATA-1能特异性结合XXIII片段内的两个GATA基序,而GATA-20不能。此外,我们还表明GATA-1也能特异性结合GAPA基因启动子中的一个顺式作用元件,该元件与GAPB基因受光协同调控。基于这些结果,我们提出光部分通过调节同一转录因子在其GATA基序上的结合来控制GAPA和GAPB基因的表达。