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串联基因扩增恢复了蓝细菌细胞色素 b 突变体中光合系统 II 的积累。

Tandem gene amplification restores photosystem II accumulation in cytochrome b mutants of cyanobacteria.

机构信息

Institute of Plant and Microbial Biology, Academia Sinica, Taipei, 11529, Taiwan.

Laboratory of Photosynthesis, Centre Algatech, Institute of Microbiology of the Czech Academy of Sciences, Třeboň, 379 01, Czech Republic.

出版信息

New Phytol. 2022 Jan;233(2):766-780. doi: 10.1111/nph.17785. Epub 2021 Oct 30.

DOI:10.1111/nph.17785
PMID:34625967
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9297868/
Abstract

Cytochrome (Cyt) b is a key component of the photosystem II complex (PSII) that is essential for its proper functioning and assembly. Site-directed mutants of the model cyanobacterium Synechocystis sp. PCC6803 with mutated heme axial ligands of Cyt b have little PSII and are therefore unable to grow photoautotrophically. Here we describe two types of Synechocystis autotrophic transformants that retained the same mutations in Cyt b but are able to accumulate PSII and grow photoautotrophically. Whole-genome sequencing revealed that all of these autotrophic transformants carried a variable number of tandem repeats (from 5 to 15) of chromosomal segments containing the psbEFLJ operon. RNA-seq analysis showed greatly increased transcript levels of the psbEFLJ operon in these autotrophic transformants. Multiple copies of the psbEFLJ operon in these transformants were only maintained during autotrophic growth, while its copy numbers gradually decreased under photoheterotrophic conditions. Two-dimensional PAGE analysis of membrane proteins revealed a strong deficiency in PSII complexes in the Cyt b mutants that was reversed in the autotrophic transformants. These results illustrate how tandem gene amplification restores PSII accumulation and photoautotrophic growth in Cyt b mutants of cyanobacteria, and may serve as an important adaptive mechanism for cyanobacterial survival.

摘要

细胞色素 b（Cyt b）是光系统 II 复合物（PSII）的关键组成部分，对其正常功能和组装至关重要。突变模型蓝藻集胞藻 PCC6803 的细胞色素 b 的血红素轴向配体的定点突变体几乎没有 PSII，因此无法进行光自养生长。在这里，我们描述了两种集胞藻能够积累 PSII 并进行光自养生长的自养转化体，它们保留了细胞色素 b 中的相同突变。全基因组测序显示，所有这些自养转化体都携带了数量可变的（5 到 15 个）含有 psbEFLJ 操纵子的染色体片段串联重复。RNA-seq 分析表明，这些自养转化体中 psbEFLJ 操纵子的转录本水平大大增加。这些转化体中的 psbEFLJ 操纵子的多个拷贝仅在自养生长期间维持，而在光异养条件下其拷贝数逐渐减少。二维 PAGE 分析膜蛋白显示，细胞色素 b 突变体中的 PSII 复合物严重不足，而在自养转化体中则得到了逆转。这些结果说明了串联基因扩增如何恢复 PSII 的积累和蓝藻细胞色素 b 突变体的光自养生长，并且可能是蓝藻生存的一种重要适应机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3d4c/9297868/4de04d775c74/NPH-233-766-g003.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3d4c/9297868/b13da95150bb/NPH-233-766-g009.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3d4c/9297868/a7fc308a2127/NPH-233-766-g006.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3d4c/9297868/c0a44493d6c3/NPH-233-766-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3d4c/9297868/f9c54f2ab9cc/NPH-233-766-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3d4c/9297868/9063db5f13c1/NPH-233-766-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3d4c/9297868/6a84c0666b16/NPH-233-766-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3d4c/9297868/b13da95150bb/NPH-233-766-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3d4c/9297868/333d68740b5b/NPH-233-766-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3d4c/9297868/4de04d775c74/NPH-233-766-g003.jpg

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