In vivo organization of the FtsZ-ring by ZapA and ZapB revealed by quantitative super-resolution microscopy.

In vivo organization of the FtsZ-ring by ZapA and ZapB revealed by quantitative super-resolution microscopy.
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DOI:
10.1111/mmi.12331
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发表时间:
2013-09
影响因子:
3.6
通讯作者:
Xiao J
Xiao J
中科院分区:
生物学2区
文献类型:
--
作者:
Buss J;Coltharp C;Huang T;Pohlmeyer C;Wang SC;Hatem C;Xiao J

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在大多数细菌细胞中,细胞分裂依赖于FtsZ蛋白的聚合,在中间细胞形成环状结构(Z环)。尽管Z环具有重要的作用,但它的分子结构仍然难以捉摸。在这项工作中,我们研究了两个FtsZ相关蛋白,ZapA和ZapB,在大肠杆菌细胞中的组装动力学和Z环结构中的作用。在缺失ZapA或ZapB的细胞中,我们观察到异常的隔膜和高动态的FtsZ结构。虽然这些FtsZ结构的细节很难在常规荧光显微镜下分辨出来,但基于单分子的超分辨成像方法光活化定位显微镜(Palm)揭示了这些FtsZ结构是由FtsZ团簇的无序排列引起的。定量分析发现,这些簇比单个FtsZ原丝更大,包含更多的分子,可能代表了Z环组装途径所固有的一种不同的聚合物物种。此外,我们发现这些簇不是由于ΔZAPA和ΔZAPB细胞中ZapB-MatP相互作用的丧失所致。我们的结果表明,ZapA和ZapB在体内的主要功能可能不是促进单个原丝的联合,而是使由多个FtsZ原丝组成的FtsZ簇排列。
In most bacterial cells, cell division is dependent on the polymerization of the FtsZ protein to form a ring-like structure (Z-ring) at the midcell. Despite its essential role, the molecular architecture of the Z-ring remains elusive. In this work we examine the roles of two FtsZ-associated proteins, ZapA and ZapB, in the assembly dynamics and structure of the Z-ring in E. coli cells. In cells deleted of zapA or zapB, we observed abnormal septa and highly dynamic FtsZ structures. While details of these FtsZ structures are difficult to discern under conventional fluorescence microscopy, single-molecule based superresolution imaging method Photoactivated Localization Microscopy (PALM) reveals that these FtsZ structures arise from disordered arrangements of FtsZ clusters. Quantitative analysis finds these clusters are larger and comprise more molecules than a single FtsZ protofilament, and likely represent a distinct polymeric species that is inherent to the assembly pathway of the Z-ring. Furthermore, we find these clusters are not due to the loss of ZapB-MatP interaction in ΔzapA and ΔzapB cells. Our results suggest that the main function of ZapA and ZapB in vivo may not be to promote the association of individual protofilaments but to align FtsZ clusters that consist of multiple FtsZ protofilaments.
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