单链DNA结合蛋白(SSB)在DNA代谢中起着至关重要的作用。SSB家族的蛋白质专门和短暂地与ssDNA结合,防止DNA双螺旋重新退火并保持基因组的完整性。同时,它们与对DNA复制至关重要的各种蛋白质相互作用和协调,重组,和修复。尽管SSB对DNA代谢至关重要,SSB家族的蛋白质长期以来一直被描述为附属物,主要是由于它们与DNA及其伙伴的动力学和机械相互作用不清楚。最近开发的单分子工具,结合生化集成技术和结构方法,增强了我们对SSB在DNA代谢过程中发挥的不同协调作用的理解。在这次审查中,我们讨论单分子检测,比如光学镊子,磁性镊子,Förster共振能量转移,以及它们的组合,增进了我们对SSB与ssDNA的结合动力学及其与其他蛋白质伴侣的相互作用的理解。我们强调了SSB蛋白通过直接调节其他蛋白的活性而发挥的中心协调作用。而不是作为附属品玩家。讨论了SSB与蛋白质伴侣相互作用的许多可能模式,它们共同提供了由SSB塑造的交互网络的更大图景。
Single-stranded DNA-binding proteins (SSBs) play vital roles in DNA metabolism. Proteins of the SSB family exclusively and transiently bind to ssDNA, preventing the DNA double helix from re-annealing and maintaining genome integrity. In the meantime, they interact and coordinate with various proteins vital for DNA replication, recombination, and repair. Although SSB is essential for DNA metabolism, proteins of the SSB family have been long described as accessory players, primarily due to their unclear dynamics and mechanistic interaction with DNA and its partners. Recently-developed single-molecule tools, together with biochemical ensemble techniques and structural methods, have enhanced our understanding of the different coordination roles that SSB plays during DNA metabolism. In this review, we discuss how single-molecule assays, such as optical tweezers, magnetic tweezers, Förster resonance energy transfer, and their combinations, have advanced our understanding of the binding dynamics of SSBs to ssDNA and their interaction with other proteins partners. We highlight the central coordination role that the SSB protein plays by directly modulating other proteins\' activities, rather than as an accessory player. Many possible modes of SSB interaction with protein partners are discussed, which together provide a bigger picture of the interaction network shaped by SSB.