Streptococcus pyogenes pSM19035 requires dynamic assembly of ATP-bound ParA and ParB on parS DNA during plasmid segregation.
Streptococcus pyogenes pSM19035 requires dynamic assembly of ATP-bound ParA and ParB on parS DNA during plasmid segregation.
复制标题
DOI：
10.1093/nar/gkn170
复制
发表时间：
2008-06
影响因子：
14.9
通讯作者：
Alonso JC
中科院分区：
文献类型：
--
作者：
Pratto F;Cicek A;Weihofen WA;Lurz R;Saenger W;Alonso JC
厚壁菌质粒pSM 19035在细胞分裂时的准确分配依赖于ATP结合和同二聚体ATP酶δ2（帕拉）的水解以及ω2（ParB）与其同源parS DNA的结合。δ2与不可水解的ATPγS形成的复合物的晶体结构分辨率为1.83 nm，揭示了一种独特的帕拉二聚体组装，它允许核苷酸交换而不需要解离成单体。在体外，在ω2和parS DNA不存在的情况下，δ2具有最小的ATP酶活性。ω2和parS DNA的化学计量比刺激δ2 ATP酶活性并介导质粒配对，而ω2：δ2高比例（4：1）时，对ATP酶活性的刺激降低，δ2聚合到DNA上。δ2 ATP酶活性的激活及其在DNA上的聚合需要ω2结合parS DNA及其N-末端的能力。体内实验表明，δ2单独与类核结合，当ω2和parS DNA存在时，δ2在类核和细胞两极之间振荡，形成螺旋状结构。我们的研究表明，ω2：δ2的摩尔比调节了（δ·ATP·Mg 2+）2在parS DNA上的聚合和解聚特性，从而控制了pSM 19035在细胞分裂前的时空分离。
The accurate partitioning of Firmicute plasmid pSM19035 at cell division depends on ATP binding and hydrolysis by homodimeric ATPase δ2 (ParA) and binding of ω2 (ParB) to its cognate parS DNA. The 1.83 Å resolution crystal structure of δ2 in a complex with non-hydrolyzable ATPγS reveals a unique ParA dimer assembly that permits nucleotide exchange without requiring dissociation into monomers. In vitro, δ2 had minimal ATPase activity in the absence of ω2 and parS DNA. However, stoichiometric amounts of ω2 and parS DNA stimulated the δ2 ATPase activity and mediated plasmid pairing, whereas at high (4:1) ω2 : δ2 ratios, stimulation of the ATPase activity was reduced and δ2 polymerized onto DNA. Stimulation of the δ2 ATPase activity and its polymerization on DNA required ability of ω2 to bind parS DNA and its N-terminus. In vivo experiments showed that δ2 alone associated with the nucleoid, and in the presence of ω2 and parS DNA, δ2 oscillated between the nucleoid and the cell poles and formed spiral-like structures. Our studies indicate that the molar ω2 : δ2 ratio regulates the polymerization properties of (δ•ATP•Mg2+)2 on and depolymerization from parS DNA, thereby controlling the temporal and spatial segregation of pSM19035 before cell division.
登录
查看更多内容
影响因子：
3.5
作者：
CEGLOWSKI, P;BOITSOV, A;ALONSO, JC
通讯作者：
ALONSO, JC
影响因子：
3.6
作者：
Ebersbach, Gitte;Ringgaard, Simon;Gerdes, Kenn
通讯作者：
Gerdes, Kenn
影响因子：
3.6
作者：
Hatano, Toshiyuki;Yamaichi, Yoshiharu;Niki, Hironori
通讯作者：
Niki, Hironori
影响因子：
10.5
作者：
Larsen, Rachel A.;Cusumano, Christina;Pogliano, Joe
通讯作者：
Pogliano, Joe
影响因子：
11.4
作者：
Barillá, D;Rosenberg, MF;Hayes, F
通讯作者：
Hayes, F