OLIGONUCLEOTIDE-DIRECTED MUTAGENESIS OF PSBB, THE GENE ENCODING CP47, EMPLOYING A DELETION MUTANT STRAIN OF THE CYANOBACTERIUM SYNECHOCYSTIS SP PCC-6803
OLIGONUCLEOTIDE-DIRECTED MUTAGENESIS OF PSBB, THE GENE ENCODING CP47, EMPLOYING A DELETION MUTANT STRAIN OF THE CYANOBACTERIUM SYNECHOCYSTIS SP PCC-6803
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DOI：
10.1007/bf00028733
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发表时间：
1991-12-01
影响因子：
5.1
通讯作者：
VERMAAS, WFJ
中科院分区：
文献类型：
--
作者：
EATONRYE, JJ;VERMAAS, WFJ
蓝藻集胞藻属的突变菌株。 PCC（巴斯德培养物保藏中心）6803 已被开发，其中 psbB（编码光系统 II (PSII) 中叶绿素 a 结合蛋白 CP47 的基因）已被删除。该缺失突变体可用于将修饰的 psbB 重新引入蓝藻中。为了研究 CP47 中大亲水区域（可能位于第五和第六跨膜区域之间的类囊体膜的腔侧）的作用，在该结构域内区域的 psbB 编码中引入了特定的删除。一个 psbB 突变导致 CP47 中的 Gly-351 缺失为 Thr-365，另一种 psbB 突变导致该蛋白中的 Arg-384 缺失为 Val-392。从 Gly-351 到 Thr-365 的缺失导致突变体 PSII 活性和光合自养生长的丧失，但 Arg-384 和 Val-392 之间的缺失保留了 PSII 活性和光合自养生长的能力。删除了 Gly-351 至 Thr-365 的突变菌株在其类囊体膜中无法组装稳定的 PSII 反应中心复合物，并且表现出 CP47 以及反应中心蛋白 D1 和 D2 水平降低。与该结构域的 Arg-384 至 Val-392 部分相反，Gly-351 和 Thr-365 之间的区域似乎对于光系统 II 的正常结构和功能至关重要。
A mutant strain of the cyanobacterium Synechocystis sp. PCC (Pasteur Culture Collection) 6803 has been developed in which psbB, the gene coding for the chlorophyl a-binding protein CP47 in Photosystem II (PSII), has been deleted. This deletion mutant can be used for the reintroduction of modified psbB into the cyanobacterium. To study the role of a large hydrophilic region in CP47, presumably located on the lumenal side of the thylakoid membrane between the fifth and sixth membrane-spanning regions, specific deletions have been introduced in psbB coding for regions within this domain. One psbB mutation leads to deletion of Gly-351 to Thr-365 in CP47, another psbB mutation was targeted towards deletion of Arg-384 to Val-392 in this protein. The deletion from Gly-351 to Thr-365 results in a loss of PSII activity and of photoautotrophic growth of the mutant, but the deletion between Arg-384 and Val-392 retains PSII activity and the ability to grow photoautotrophically. The mutant strain with the deletion from Gly-351 to Thr-365 does not assemble a stable PSII reaction center complex in its thylakoid membranes, and exhibits diminished levels of CP47 and of the reaction center proteins D1 and D2. In contrast to the Arg-384 to Val-392 portion of this domain, the region between Gly-351 and Thr-365 appears essential for the normal structure and function of photosystem II.