Regulation of DNA replication in B.subtilis by YabA

描述(申请人提供)：用于控制革兰氏阳性细菌复制启动的时间和频率的机制(S)尚不清楚。Yaba是枯草杆菌复制起始的负调控因子，在其他革兰氏阳性菌中也是保守的。Yaba执行其调控功能的机制尚不清楚。我发现Yaba与染色体复制起点(ORIC)有关，并在复制解旋酶加载后的一个步骤中递送到那里。我建议进行一系列实验，以确定将八巴与奥利克联系起来所必需的因素(S)。我还建议进行实验，以确定Yaba的亚细胞位置及其与染色体的关系是否在复制周期中发生变化。这些实验应该为Yaba在抑制复制启动中作用的时机提供洞察力。最后，我提出了几个体内和体外实验，以确定Yaba抑制复制启动的机制。

DESCRIPTION (provided by applicant): The mechanism(s) used to control the timing and frequency of replication initiation in Gram positive bacteria are unknown. YabA is a negative regulator of replication initiation in B. subtilis, and is conserved in other Gram positive bacteria. The mechanism by which YabA carries out its regulatory function is not yet understood. I found that YabA associates with the chromosomal origin of replication (oriC), and is delivered there at a step following the loading of the replicative helicase. I propose a series of experiments to identify the factor(s) necessary for association of YabA with oriC. I also propose experiments to determine if the subcellular location of YabA and its association with the chromosome changes during the replication cycle. These experiments should provide insights into the timing of YabA's function in inhibition of replication initiation. Finally, I propose several in vivo and in vitro experiments to determine the mechanism of inhibition of replication initiation by YabA.