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FlhF是一种信号识别颗粒样GTP酶,参与蜡样芽孢杆菌中鞭毛排列、运动行为和蛋白质分泌的调控。

FlhF, a signal recognition particle-like GTPase, is involved in the regulation of flagellar arrangement, motility behaviour and protein secretion in Bacillus cereus.

作者信息

Salvetti Sara, Ghelardi Emilia, Celandroni Francesco, Ceragioli Mara, Giannessi Francesco, Senesi Sonia

机构信息

Dipartimento di Patologia Sperimentale, Biotecnologie Mediche, Infettivologia ed Epidemiologia, Università di Pisa, via San Zeno 37, 56127 Pisa, Italy.

Dipartimento di Biologia, Sezione di Microbiologia, Università di Pisa, via San Zeno 35, 56127 Pisa, Italy.

出版信息

Microbiology (Reading). 2007 Aug;153(Pt 8):2541-2552. doi: 10.1099/mic.0.2006/005553-0.

Abstract

Flagellar arrangement is a highly conserved feature within bacterial species. However, only a few genes regulating cell flagellation have been described in polar flagellate bacteria. This report demonstrates that the arrangement of flagella in the peritrichous flagellate Bacillus cereus is controlled by flhF. Disruption of flhF in B. cereus led to a reduction in the number of flagella from 10-12 to 1-3 filaments per cell in the insertion mutant MP06. Moreover, compared to the parental strain, MP06 exhibited: (i) shorter smooth swimming phases, causing reduced swimming motility but not affecting chemotaxis; (ii) complete inhibition of swarming motility, as differentiated swarm cells were never detected; (iii) an increased amount of extracellular proteins; and (iv) differential export of virulence determinants, such as haemolysin BL (HBL), phosphatidylcholine-preferring phospholipase C (PC-PLC) and non-haemolytic enterotoxin (NHE). Introduction of a plasmid harbouring flhF (pDGflhF) into MP06 completely restored the wild-type phenotype in the trans-complemented strain MP07. B. cereus flhF was found to constitute a monocistronic transcriptional unit and its overexpression did not produce abnormal features in the wild-type background. Characterization of a B. cereus mutant (MP05) carrying a partial flhF deletion indicated that the last C-terminal domain of FlhF is involved in protein export while not required for flagellar arrangement and motility behaviour. Taken together, these data suggest that B. cereus FlhF is a promising candidate for connecting diverse cellular functions, such as flagellar arrangement, motility behaviour, pattern of protein secretion and virulence phenotype.

摘要

鞭毛排列是细菌物种内一个高度保守的特征。然而,在极生鞭毛细菌中,仅有少数调节细胞鞭毛形成的基因被描述过。本报告表明,周生鞭毛的蜡样芽孢杆菌中鞭毛的排列受flhF控制。蜡样芽孢杆菌中flhF的缺失导致插入突变体MP06中每个细胞的鞭毛数量从10 - 12根减少到1 - 3根丝状体。此外,与亲本菌株相比,MP06表现出:(i) 平滑游动阶段更短,导致游动运动性降低,但不影响趋化性;(ii) 群体运动完全受到抑制,因为从未检测到分化的群体细胞;(iii) 细胞外蛋白量增加;以及(iv) 毒力决定因子的差异输出,如溶血素BL(HBL)、偏好磷脂酰胆碱的磷脂酶C(PC-PLC)和非溶血肠毒素(NHE)。将携带flhF的质粒(pDGflhF)导入MP06,在反式互补菌株MP07中完全恢复了野生型表型。发现蜡样芽孢杆菌flhF构成一个单顺反子转录单元,其在野生型背景下的过表达未产生异常特征。对携带部分flhF缺失的蜡样芽孢杆菌突变体(MP05)的表征表明,FlhF的最后一个C末端结构域参与蛋白质输出,而对鞭毛排列和运动行为不是必需的。综上所述,这些数据表明蜡样芽孢杆菌FlhF是连接多种细胞功能(如鞭毛排列、运动行为、蛋白质分泌模式和毒力表型)的一个有前景的候选者。

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