Brendler T, Sawitzke J, Sergueev K, Austin S
Gene Regulation and Chromosome Biology Laboratory, Division of Basic Sciences, NCI-Frederick Cancer Research and Development Center, Frederick, MD 21702-1201, USA.
EMBO J. 2000 Nov 15;19(22):6249-58. doi: 10.1093/emboj/19.22.6249.
SeqA is an Escherichia coli DNA-binding protein that acts at replication origins and controls DNA replication. However, binding is not exclusive to origins. Many fragments containing two or more hemi-methylated GATC sequences bind efficiently. Binding was optimal when two such sequences were closely apposed or up to 31 bases apart on the same face of the DNA helix. Binding studies suggest that neighboring bound proteins contact each other to form a complex with the intervening DNA looped out. There are many potential binding sites distributed around the E.coli chromosome. As replication produces a transient wave of hemi-methylation, tracts of SeqA binding are likely to associate with each fork as replication progresses. The number and positions of green fluorescent protein-SeqA foci seen in living cells suggest that they correspond to these tracts, and that the forks are tethered to planes of cell division. SeqA may help to tether the forks or to organize newly replicated DNA into a structure that aids DNA to segregate away from the replication machinery.
SeqA是一种大肠杆菌DNA结合蛋白,作用于复制起点并控制DNA复制。然而,其结合并不局限于起点。许多含有两个或更多半甲基化GATC序列的片段能有效结合。当两个这样的序列紧密相邻或在DNA螺旋同一面上相隔多达31个碱基时,结合最为理想。结合研究表明,相邻的结合蛋白相互接触,形成一个中间DNA呈环状突出的复合物。大肠杆菌染色体周围分布着许多潜在的结合位点。随着复制产生半甲基化的瞬时波,随着复制的进行,SeqA结合区域可能会与每个复制叉相关联。在活细胞中观察到的绿色荧光蛋白-SeqA焦点的数量和位置表明,它们与这些区域相对应,并且复制叉与细胞分裂平面相连。SeqA可能有助于固定复制叉,或将新复制的DNA组织成一种有助于DNA与复制机器分离的结构。