Bustos S A, Golden S S
Department of Biology, Texas A & M University, College Station 77843-3258.
J Bacteriol. 1991 Dec;173(23):7525-33. doi: 10.1128/jb.173.23.7525-7533.1991.
The psbDI and psbDII genes in Synechococcus sp. strain PCC 7942 encode the D2 polypeptide, an essential component of the photosystem II reaction center. Previous studies have demonstrated that transcripts from psbDII, but not psbDI, increase in response to high light intensity. Soluble proteins from Synechococcus cells shifted to high light were found to have affinity for DNA sequences upstream from the psbDII coding region. DNA mobility-shift and copper-phenanthroline footprinting assays of a 258-bp fragment revealed three distinct DNA-protein complexes that mapped to the untranslated leader region between +11 and +84. Deletion of the upstream flanking region to -42 had no effect on the expression of a psbDII-lacZ reporter gene or its induction by light, whereas a promoterless construct supported only minimal background levels of beta-galactosidase. A 4-bp deletion within the first protected region of the footprint decreased the beta-galactosidase activity to approximately 2% of that of the undeleted control, but gene expression remained responsive to light. Deletion of the three protected regions completely abolished both gene expression and light induction. These results suggest that the psbDII gene requires elements within the untranslated leader region for efficient gene expression, one of which may be involved in regulation by light.
聚球藻属(Synechococcus)PCC 7942菌株中的psbDI和psbDII基因编码D2多肽,它是光系统II反应中心的一个必需组分。先前的研究表明,响应高光强度时,psbDII而非psbDI的转录本会增加。研究发现,转移至高光条件下的聚球藻细胞中的可溶性蛋白对psbDII编码区上游的DNA序列具有亲和力。对一个258 bp片段进行的DNA迁移率变动分析和铜菲咯啉足迹分析揭示出三种不同的DNA-蛋白质复合物,它们定位于+11至+84之间的非翻译前导区。将上游侧翼区域缺失至-42对psbDII - lacZ报告基因的表达或其光诱导没有影响,而一个无启动子的构建体仅支持极低水平的β-半乳糖苷酶背景表达。足迹分析的第一个受保护区域内的一个4 bp缺失使β-半乳糖苷酶活性降至未缺失对照的约2%,但基因表达仍对光有响应。三个受保护区域的缺失完全消除了基因表达和光诱导。这些结果表明,psbDII基因需要非翻译前导区内的元件来实现高效基因表达,其中一个元件可能参与光调节。