Transient non-specific DNA binding dominates the target search of bacterial DNA-binding proteins

Transient non-specific DNA binding dominates the target search of bacterial DNA-binding proteins
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瞬时非特异性 DNA 结合主导细菌 DNA 结合蛋白的靶标搜索

DOI:
10.1101/2020.08.13.249771
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发表时间:
2020
期刊:
--
影响因子:
--
通讯作者:
Stracy M
Stracy M
中科院分区:
--
文献类型:
--
作者:
Stracy M

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尽管它们具有不同的生化特性和功能,但所有DNA结合蛋白都具有在大量过量的非靶DNA中准确定位其靶位点的能力。为了确定目标搜索的通用机制,我们使用了活大肠杆菌中11种不同DNA结合蛋白的单分子跟踪。这些蛋白质在目标搜索期间的移动性由DNA相互作用决定,而不是由它们的分子量决定。通过产生缺乏所有染色体DNA的细胞,我们发现类核不是蛋白质扩散的物理屏障,而是通过频繁的短暂DNA相互作用显着减缓DNA结合蛋白的运动。代表性的DNA结合蛋白(无论其大小、浓度或功能如何)花费其大部分(58%-99%)的搜索时间与DNA结合,并且在任何时候占据多达30%的染色体DNA。染色体拥挤可能对所有DNA结合蛋白的功能具有重要意义。
Despite their diverse biochemical characteristics and functions, all DNA-binding proteins share the ability to accurately locate their target sites among the vast excess of non-target DNA. Toward identifying universal mechanisms of the target search, we used single-molecule tracking of 11 diverse DNA-binding proteins in livingEscherichia coli. The mobility of these proteins during the target search was dictated by DNA interactions rather than by their molecular weights. By generating cells devoid of all chromosomal DNA, we discovered that the nucleoid is not a physical barrier for protein diffusion but significantly slows the motion of DNA-binding proteins through frequent short-lived DNA interactions. The representative DNA-binding proteins (irrespective of their size, concentration, or function) spend the majority (58%–99%) of their search time bound to DNA and occupy as much as ∼30% of the chromosomal DNA at any time. Chromosome crowding likely has important implications for the function of all DNA-binding proteins.
拓扑异构酶 IV 在大肠杆菌染色体分离中的定位和作用由 SMC 复合体 MukBEF 协调
DOI: --
发表时间: 2015
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