Dissection of the region of Pseudomonas aeruginosa ParA that is important for dimerization and interactions with its partner ParB.
Dissection of the region of Pseudomonas aeruginosa ParA that is important for dimerization and interactions with its partner ParB.
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DOI：
10.1099/mic.0.081216-0
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发表时间：
2014-11
期刊：
影响因子：
--
通讯作者：
Jagura-Burdzy G
中科院分区：
文献类型：
--
作者：
Bartosik AA;Glabski K;Jecz P;Lasocki K;Mikosa M;Plochocka D;Thomas CM;Jagura-Burdzy G
铜绿假单胞菌 ParA 属于 Walker 型 ATP 酶的一个大亚家族，在细菌中充当分配蛋白。 ParA 具有自关联和与其伙伴 ParB 交互的能力。对缺失突变体的分析确定了参与二聚化以及与 ParB 相互作用的蛋白质部分。在这里，在 E67 和 L85 之间的区域创建了一组 ParA 丙氨酸取代突变体，并在体内和体外进行了分析。所有二聚化受损的突变体（M74、H79、Y82 和 L84 位点的取代）在与 ParB 的相互作用中也存在缺陷，这表明 ParA-ParB 相互作用取决于 ParA 二聚化的能力。在 E67、C68、L70、E72、F76、Q83 和 L85 位点具有丙氨酸取代的突变体的二聚化并未受到损害，但与 ParB 的相互作用存在缺陷。二聚化界面部分与假发夹重叠，参与与 ParB 的相互作用。体外测试的ParA突变衍生物显示ATP酶活性没有缺陷。通过同源基因交换将两个 parA 等位基因（parA84，其产物既不能自相互作用，也不能与 ParB 相互作用，以及 parA67，其产物在与 ParB 的相互作用中受到损害，但在二聚化中不受损害）被引入铜绿假单胞菌染色体。两种突变体均表现出 ParB 焦点分离缺陷，但程度不同。只有 PAO1161 parA84 在染色体分离、生长速率和运动性方面明显受损，类似于 parA 缺失突变体。
Pseudomonas aeruginosa ParA belongs to a large subfamily of Walker-type ATPases acting as partitioning proteins in bacteria. ParA has the ability to both self-associate and interact with its partner ParB. Analysis of the deletion mutants defined the part of the protein involved in dimerization and interactions with ParB. Here, a set of ParA alanine substitution mutants in the region between E67 and L85 was created and analysed in vivo and in vitro. All mutants impaired in dimerization (substitutions at positions M74, H79, Y82 and L84) were also defective in interactions with ParB, suggesting that ParA–ParB interactions depend on the ability of ParA to dimerize. Mutants with alanine substitutions at positions E67, C68, L70, E72, F76, Q83 and L85 were not impaired in dimerization, but were defective in interactions with ParB. The dimerization interface partly overlapped the pseudo-hairpin, involved in interactions with ParB. ParA mutant derivatives tested in vitro showed no defects in ATPase activity. Two parA alleles (parA84, whose product can neither self-interact nor interact with ParB, and parA67, whose product is impaired in interactions with ParB, but not in dimerization) were introduced into the P. aeruginosa chromosome by homologous gene exchange. Both mutants showed defective separation of ParB foci, but to different extents. Only PAO1161 parA84 was visibly impaired in terms of chromosome segregation, growth rate and motility, similar to a parA-null mutant.
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影响因子：
3.2
作者：
Kusiak, Magdalena;Gapczynska, Anna;Jagura-Burdzy, Grazyna
通讯作者：
Jagura-Burdzy, Grazyna
影响因子：
11.4
作者：
Bouet, JY;Funnell, BE
通讯作者：
Funnell, BE
影响因子：
4.5
作者：
Ah-Seng, Yoan;Rech, Jerome;Bouet, Jean-Yves
通讯作者：
Bouet, Jean-Yves
影响因子：
2.8
作者：
Dmowski, Michal;Jagura-Burdzy, Grazyna
通讯作者：
Jagura-Burdzy, Grazyna
影响因子：
3.7
作者：
Bartosik AA;Glabski K;Jecz P;Mikulska S;Fogtman A;Koblowska M;Jagura-Burdzy G
通讯作者：
Jagura-Burdzy G