Structure-function studies reveal ComEA contains an oligomerization domain essential for transformation in gram-positive bacteria.
Structure-function studies reveal ComEA contains an oligomerization domain essential for transformation in gram-positive bacteria.
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DOI：
10.1038/s41467-022-35129-0
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发表时间：
2022-12-13
影响因子：
16.6
通讯作者：
Neiditch, Matthew B.
中科院分区：
文献类型：
--
作者：
Ahmed, Ishtiyaq;Hahn, Jeanette;Henrickson, Amy;Khaja, Faisal Tarique;Demeler, Borries;Dubnau, David;Neiditch, Matthew B.
细菌转化的一个重要步骤是将DNA摄取到周质中，穿过革兰氏阳性细菌的厚肽聚糖细胞壁，或革兰氏阴性细菌的外膜和薄肽聚糖层。ComEA是一种在可转化细菌中广泛保守的DNA结合蛋白，是该摄取步骤所必需的。在这里，我们确定了两种革兰氏阳性菌（枯草芽孢杆菌和嗜热脂肪土芽孢杆菌）的ComEA的X射线晶体结构，确定了革兰氏阴性菌中不存在的结构域。X射线晶体学，遗传学和分析超离心（AUC）分析表明，这个域驱动ComEA寡聚化，我们表明这是必要的转换。我们使用多波长AUC（MW-AUC）来表征DNA和ComEA DNA结合结构域之间的相互作用。最后，我们提出了一个模型的ComEA DNA结合结构域与DNA的相互作用，表明ComEA寡聚化可以提供一个拉力，驱动DNA吸收的革兰氏阳性菌的厚细胞壁。ComEA是细菌转化过程中DNA摄取所需的DNA结合蛋白。在这里，Ahmed等人确定了来自革兰氏阳性细菌的ComEA的X射线晶体结构，鉴定了革兰氏阴性细菌中不存在的结构域，并驱动ComEA寡聚化，这是转化所需的。
An essential step in bacterial transformation is the uptake of DNA into the periplasm, across the thick peptidoglycan cell wall of Gram-positive bacteria, or the outer membrane and thin peptidoglycan layer of Gram-negative bacteria. ComEA, a DNA-binding protein widely conserved in transformable bacteria, is required for this uptake step. Here we determine X-ray crystal structures of ComEA from two Gram-positive species, Bacillus subtilis and Geobacillus stearothermophilus, identifying a domain that is absent in Gram-negative bacteria. X-ray crystallographic, genetic, and analytical ultracentrifugation (AUC) analyses reveal that this domain drives ComEA oligomerization, which we show is required for transformation. We use multi-wavelength AUC (MW-AUC) to characterize the interaction between DNA and the ComEA DNA-binding domain. Finally, we present a model for the interaction of the ComEA DNA-binding domain with DNA, suggesting that ComEA oligomerization may provide a pulling force that drives DNA uptake across the thick cell walls of Gram-positive bacteria. ComEA is a DNA-binding protein required for DNA uptake during bacterial transformation. Here, Ahmed et al. determine X-ray crystal structures of ComEA from Gram-positive bacteria, identifying a domain that is absent in Gram-negative bacteria and drives ComEA oligomerization, which is required for transformation.
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影响因子：
3.4
作者：
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通讯作者：
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影响因子：
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作者：
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Maier B
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K