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转录组和缺失突变分析表明,在碳饥饿条件下,玫瑰色杆菌中一种 RpoH 家族 σ 因子对于光系统的产生是必需的。

Transcriptome and Deletion Mutant Analyses Revealed that an RpoH Family Sigma Factor Is Essential for Photosystem Production in Roseateles depolymerans under Carbon Starvation.

机构信息

Bio-Analytical Research Group, Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST).

Photosynthetic Microbial Consortia Laboratory, Department of Biological Sciences, Graduate School of Science, Tokyo Metropolitan University.

出版信息

Microbes Environ. 2023;38(1). doi: 10.1264/jsme2.ME22072.

Abstract

Roseateles depolymerans is an obligately aerobic bacterium that produces a photosynthetic apparatus only under the scarcity of carbon substrates. We herein examined changes in the transcriptomes of R. depolymerans cells to clarify the expression of photosynthesis genes and their upstream regulatory factors under carbon starvation. Transcriptomes 0, 1, and 6‍ ‍h after the depletion of a carbon substrate indicated that transcripts showing the greatest variations (a 500-fold increase [6 h/0 h]) were light-harvesting proteins (PufA and PufB). Moreover, loci with more than 50-fold increases (6 h/0‍ ‍h) were fully related to the photosynthetic gene cluster. Among 13 sigma factor genes, the transcripts of a sigma 70 family sigma factor related to RpoH (SP70) increased along photosynthesis genes under starvation; therefore, a knockout experiment of SP70 was performed. ΔSP70 mutants were found to lack photosynthetic pigments (carotenoids and bacteriochlo-rophyll a) regardless of carbon starvation. We also examined the effects of heat stress on ΔSP70 mutants, and found that SP70 was also related to heat stress tolerance, similar to other RpoH sigma factors (while heat stress did not trigger photosystem production). The deficient accumulation of photosynthetic pigments and the heat stress tolerance of ΔSP70 mutants were both complemented by the introduction of an intact SP70 gene. Furthermore, the transcription of photosynthetic gene operons (puf, puh, and bch) was markedly reduced in the ΔSP70 mutant. The RpoH homologue SP70 was concluded to be a sigma factor that is essential for the transcription of photosynthetic gene operons in R. depolymerans.

摘要

玫瑰色射电杆菌是一种严格需氧细菌,只有在碳源底物匮乏的情况下才会产生光合器官。我们在此检查了 R. depolymerans 细胞的转录组变化,以阐明在碳饥饿下光合作用基因及其上游调控因子的表达情况。在耗尽碳底物后的 0、1 和 6 小时的转录组分析表明,转录本变化最大(6 h/0 h 时增加了 500 倍)的是光捕获蛋白(PufA 和 PufB)。此外,表达水平增加了 50 倍以上(6 h/0 h)的基因座与光合基因簇完全相关。在 13 个 sigma 因子基因中,与 RpoH 相关的 sigma 70 家族 sigma 因子(SP70)的转录本在饥饿时随着光合作用基因的增加而增加;因此,进行了 SP70 的敲除实验。ΔSP70 突变体缺乏光合色素(类胡萝卜素和菌绿素 a),无论是否存在碳饥饿。我们还研究了热应激对 ΔSP70 突变体的影响,发现 SP70 也与热应激耐受性有关,与其他 RpoH sigma 因子相似(而热应激不会触发光合系统的产生)。ΔSP70 突变体中光合色素的积累不足和热应激耐受性都可以通过引入完整的 SP70 基因得到弥补。此外,ΔSP70 突变体中光合基因操纵子(puf、puh 和 bch)的转录明显减少。得出结论,RpoH 同源物 SP70 是玫瑰色射电杆菌光合基因操纵子转录所必需的 sigma 因子。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ce57/10037100/d931d8542890/38_22072-g001.jpg

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