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RRNPP-type quorum-sensing systems regulate solvent formation, sporulation and cell motility in Clostridium saccharoperbutylacetonicum.
Biotechnology for Biofuels ( IF 6.3 ) Pub Date : 2020-05-08 , DOI: 10.1186/s13068-020-01723-x
Jun Feng 1 , Wenming Zong 1, 2 , Pixiang Wang 1 , Zhong-Tian Zhang 1 , Yanyan Gu 1 , Mark Dougherty 1 , Ilya Borovok 3 , Yi Wang 1, 4
Affiliation  

Background Clostridium saccharoperbutylacetonicum N1-4 (HMT) is a strictly anaerobic, spore-forming Gram-positive bacterium capable of hyper-butanol production through the well-known acetone-butanol-ethanol fermentation process. Recently, five putative RRNPP-type QSSs (here designated as QSS1 to QSS5) were predicted in this bacterial strain, each of which comprises a putative RRNPP-type regulator (QssR1 to QssR5) and a cognate signaling peptide precursor (QssP1 to QssP5). In addition, both proteins are encoded by the same operon. The functions of these multiple RRNPP-type QSSs are unknown. Results To elucidate the function of multiple RRNPP-type QSSs as related to cell metabolism and solvent production in N1-4 (HMT), we constructed qssR-deficient mutants ΔR1, ΔR2, ΔR3 and ΔR5 through gene deletion using CRISPR-Cas9 and N1-4-dcas9-R4 (with the QssR4 expression suppressed using CRISPR-dCas9). We also constructed complementation strains by overexpressing the corresponding regulator gene. Based on systematic characterization, results indicate that QSS1, QSS2, QSS3, and QSS5 positively regulate the sol operon expression and thus solvent production, but they likely negatively regulate cell motility. Consequently, QSS4 might not directly regulate solvent production, but positively affect cell migration. In addition, QSS3 and QSS5 appear to positively regulate sporulation efficiency. Conclusions Our study provides the first insights into the roles of multiple RRNPP-type QSSs of C. saccharoperbutylacetonicum for the regulation of solvent production, cell motility, and sporulation. Results of this study expand our knowledge of how multiple paralogous QSSs are involved in the regulation of essential bacterial metabolism pathways.

中文翻译:

RRNPP 型群体感应系统调节梭状芽孢杆菌中的溶剂形成、孢子形成和细胞运动。

背景 糖过丁基丙酮梭菌 N1-4 (HMT) 是一种严格厌氧、形成孢子的革兰氏阳性细菌,能够通过众所周知的丙酮-丁醇-乙醇发酵过程生产超丁醇。最近,在该菌株中预测了五种推定的 RRNPP 型 QSS(此处指定为 QSS1 至 QSS5),每个都包含推定的 RRNPP 型调节剂(QssR1 至 QssR5)和同源信号肽前体(QssP1 至 QssP5)。此外,两种蛋白质都由相同的操纵子编码。这些多个 RRNPP 型 QSS 的功能是未知的。结果 为了阐明多种 RRNPP 型 QSS 在 N1-4 (HMT) 中与细胞代谢和溶剂产生相关的功能,我们构建了 qssR 缺陷突变体 ΔR1、ΔR2、ΔR3 和 ΔR5 通过使用 CRISPR-Cas9 和 N1-4-dcas9-R4 的基因缺失(使用 CRISPR-dCas9 抑制 QssR4 表达)。我们还通过过表达相应的调节基因来构建互补菌株。基于系统表征,结果表明 QSS1、QSS2、QSS3 和 QSS5 正调节溶胶操纵子的表达,从而调节溶剂的产生,但它们可能负调节细胞运动。因此,QSS4 可能不会直接调节溶剂的产生,但会对细胞迁移产生积极影响。此外,QSS3 和 QSS5 似乎正向调节孢子形成效率。结论 我们的研究首次揭示了糖化丙酮梭菌的多种 RRNPP 型 QSS 在调节溶剂产生、细胞运动和孢子形成方面的作用。
更新日期:2020-05-08
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