Bacteriophage SP01 Gene Product 56 Inhibits Bacillus subtilis Cell Division by Interacting with FtsL and Disrupting Pbp2B and FtsW Recruitment
Bacteriophage SP01 Gene Product 56 Inhibits Bacillus subtilis Cell Division by Interacting with FtsL and Disrupting Pbp2B and FtsW Recruitment
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DOI：
10.1128/jb.00463-20
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发表时间：
2021-01-01
影响因子：
3.2
通讯作者：
Haeusser, Daniel P.
中科院分区：
文献类型：
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作者：
Bhambhani, Amit;Iadicicco, Isabella;Haeusser, Daniel P.
先前的工作鉴定了由裂解性噬菌体SP01编码的基因产物56（gp56），其负责在其感染期间抑制枯草芽孢杆菌细胞分裂。在大多数细菌中，必需的微管蛋白样蛋白FtsZ在胞质分裂的新生位点组装成环形结构决定了分裂的时间和位置。该FtsZ环作为将其他蛋白质募集到允许膜收缩和间隔细胞壁合成的成熟分裂能力结构中的支架。在这里，我们表明预测的9.3 kDa的SP01 gp56的表达抑制了B的后期阶段。枯草芽孢杆菌细胞分裂而不改变FtsZ环组装。绿色荧光蛋白标记的gp56定位于分裂位点的膜上。虽然它的定位不干扰早期分裂蛋白的募集，但gp56干扰晚期分裂蛋白的募集，包括Pbp2b和FtsW。特定分裂成分缺失或耗尽的细胞成像和双杂交分析表明，gp56的定位和活性取决于其与FtsL的相互作用。总之，这些数据支持一个模型，其中gp56与分裂机制的中心部分相互作用，以破坏参与隔细胞壁synthesization.IMPORTANCE研究在过去的几十年中已经确定了噬菌体编码的因子，干扰宿主细胞的形状或胞质分裂在病毒感染。迄今为止已经研究的引起细胞抑制的噬菌体因子都通过靶向FtsZ起作用，FtsZ是保守的原核微管蛋白同系物，其在大多数细菌和一些古细菌中构成细胞动力学环。然而，包括枯草芽孢杆菌噬菌体SP01的gp56在内的几种抑制胞质分裂的噬菌体因子的机制仍未探索。在这里，我们表明，不同于其他已发表的例子，噬菌体抑制胞质分裂，gp56块B。枯草芽孢杆菌细胞分裂而不靶向FtsZ。相反，它利用组装的FtsZ细胞动力学环定位于分裂机构，并阻止间隔细胞壁合成所需的蛋白质的募集。
Previous work identified gene product 56 (gp56), encoded by the lytic bacteriophage SP01, as being responsible for inhibition of Bacillus subtilis cell division during its infection. Assembly of the essential tubulin-like protein FtsZ into a ring-shaped structure at the nascent site of cytokinesis determines the timing and position of division in most bacteria. This FtsZ ring serves as a scaffold for recruitment of other proteins into a mature division-competent structure permitting membrane constriction and septal cell wall synthesis. Here, we show that expression of the predicted 9.3-kDa gp56 of SP01 inhibits later stages of B. subtilis cell division without altering FtsZ ring assembly. Green fluorescent protein-tagged gp56 localizes to the membrane at the site of division. While its localization does not interfere with recruitment of early division proteins, gp56 interferes with the recruitment of late division proteins, including Pbp2b and FtsW. Imaging of cells with specific division components deleted or depleted and two-hybrid analyses suggest that gp56 localization and activity depend on its interaction with FtsL. Together, these data support a model in which gp56 interacts with a central part of the division machinery to disrupt late recruitment of the division proteins involved in septal cell wall synthesis.IMPORTANCE Studies over the past decades have identified bacteriophage-encoded factors that interfere with host cell shape or cytokinesis during viral infection. The phage factors causing cell filamentation that have been investigated to date all act by targeting FtsZ, the conserved prokaryotic tubulin homolog that composes the cytokinetic ring in most bacteria and some groups of archaea. However, the mechanisms of several phage factors that inhibit cytokinesis, including gp56 of bacteriophage SP01 of Bacillus subtilis, remain unexplored. Here, we show that, unlike other published examples of phage inhibition of cytokinesis, gp56 blocks B. subtilis cell division without targeting FtsZ. Rather, it utilizes the assembled FtsZ cytokinetic ring to localize to the division machinery and to block recruitment of proteins needed for septal cell wall synthesis.