Henckes G, Harper F, Levine A, Vannier F, Séror S J
Institut de Génétique et Microbiologie, Centre National de la Recherche Scientifique 1354, Université Paris, Orsay, France.
Proc Natl Acad Sci U S A. 1989 Nov;86(22):8660-4. doi: 10.1073/pnas.86.22.8660.
When the Bacillus subtilis dnaB37 mutant, defective in initiation, is returned to permissive temperature after accumulation of initiation proteins at 45 degrees C, we have shown, by extensive DNA.DNA hybridization analysis, that the origin region is replicated in excess (approximately 2-fold). However, this replication is limited to a region of about 120-175 kilobases on either side of the origin. This has been confirmed by autoradiographic analysis of the overreplicated region. During the second round of synchronized replication at 30 degrees C, replication in fact appears to resume from the stalled forks on either side of the origin. We propose that in B. subtilis, in addition to a first level of control at the origin, a second level of control exists downstream of the origin in order to limit overreplication of the chromosome. These two controls might normally be tightly coupled. We suggest that the second level of control is exerted through the reversible inhibition of replisome movement at specific regions on either side of the origin.
当在45℃积累起始蛋白后,将起始存在缺陷的枯草芽孢杆菌dnaB37突变体恢复到允许温度时,我们通过广泛的DNA-DNA杂交分析表明,起始区域会过度复制(约2倍)。然而,这种复制仅限于起始点两侧约120-175千碱基的区域。这已通过对过度复制区域的放射自显影分析得到证实。在30℃第二轮同步复制期间,复制实际上似乎是从起始点两侧停滞的叉处重新开始的。我们提出,在枯草芽孢杆菌中,除了在起始点存在第一级控制外,在起始点下游还存在第二级控制,以限制染色体的过度复制。这两种控制通常可能紧密耦合。我们认为,第二级控制是通过对起始点两侧特定区域的复制体移动进行可逆抑制来实现的。