Kim Eun Sook, Bae Hee-Won, Cho You-Hee
Department of Pharmacy, College of Pharmacy and Institute of Pharmaceutical Sciences, CHA University, Gyeonggi-do, South Korea.
Front Microbiol. 2018 Feb 16;9:247. doi: 10.3389/fmicb.2018.00247. eCollection 2018.
The host range of a phage is determined primarily by phage-receptor interaction. Here, we profiled the host range of an RNA leviphage, PP7 that requires functional type IV pilus (TFP) in order to enter into its host bacterium, . Out of 25 twitching-proficient strains, 4 with group I pilin and 7 with group III pilin displayed PP7-resistance. The remaining 14 possessed group II pilin, which included 10 PP7-sensitive and 4 PP7-resistant strains, suggesting that only the strains with TFP consisted of a subset of group II (hence, group IIa) pilin were susceptible to PP7. The co-expression of the PAO1 (group IIa) pilin rendered all the strains susceptible to PP7, with the exception of the 4 strains with group I pilin. Moreover, the expression of PA14 (group III) and PAK (group IIb) pilin in the PAO1 mutant restored the twitching motility but not the PP7-suceptibility. Site-directed and random mutation analyses of PAO1 pilin enabled us to identify a pilin mutant (G96S) that is fully functional but resistant to PP7 infection. This is due to the lack of any phage-receptor interactions, suggesting the structural properties of the β1-β2 loop in the variable region 2 of the group II pilin might be involved in PP7 infection.
噬菌体的宿主范围主要由噬菌体-受体相互作用决定。在此,我们分析了一种RNA轻噬菌体PP7的宿主范围,该噬菌体需要功能性IV型菌毛(TFP)才能进入其宿主细菌。在25株具有抽动能力的菌株中,4株具有I型菌毛蛋白,7株具有III型菌毛蛋白,表现出对PP7的抗性。其余14株具有II型菌毛蛋白,其中包括10株对PP7敏感和4株对PP7抗性的菌株,这表明只有具有TFP且由II型(因此为IIa组)菌毛蛋白子集组成的菌株对PP7敏感。PAO1(IIa组)菌毛蛋白的共表达使所有菌株对PP7敏感,但4株具有I型菌毛蛋白的菌株除外。此外,PA14(III组)和PAK(IIb组)菌毛蛋白在PAO1突变体中的表达恢复了抽动运动性,但未恢复对PP7的敏感性。对PAO1菌毛蛋白进行定点和随机突变分析,使我们能够鉴定出一种完全功能性但对PP7感染具有抗性的菌毛蛋白突变体(G96S)。这是由于缺乏任何噬菌体-受体相互作用,表明II型菌毛蛋白可变区2中β1-β2环的结构特性可能参与了PP7感染。