Tafvizi Anahita, Huang Fang, Leith Jason S, Fersht Alan R, Mirny Leonid A, van Oijen Antoine M
Harvard-Massachusetts Institute of Technology, Division of Health Sciences and Technology, Cambridge, Massachusetts, USA.
Biophys J. 2008 Jul;95(1):L01-3. doi: 10.1529/biophysj.108.134122. Epub 2008 Apr 18.
The p53 protein, a transcription factor of key importance in tumorigenesis, is suggested to diffuse one-dimensionally along DNA via its C-terminal domain, a process that is proposed to regulate gene activation both positively and negatively. There has been no direct observation of p53 moving along DNA, however, and little is known about the mechanism and rate of its translocation. Here, we use single-molecule techniques to visualize, in real time, the one-dimensional diffusion of p53 along DNA. The one-dimensional diffusion coefficient is measured to be close to the theoretical limit, indicative of movement along a free energy landscape with low activation barriers. We further investigate the mechanism of translocation and determine that p53 is capable of sliding--moving along DNA while in continuous contact with the duplex, rather than through a series of hops between nearby bases.
p53蛋白是肿瘤发生过程中至关重要的转录因子,有人提出它通过其C末端结构域沿DNA进行一维扩散,这一过程被认为可正负调节基因激活。然而,尚未有直接观察到p53沿DNA移动的情况,其转位机制和速率也知之甚少。在此,我们使用单分子技术实时可视化p53沿DNA的一维扩散。测得的一维扩散系数接近理论极限,表明其沿着具有低活化能垒的自由能景观移动。我们进一步研究了转位机制,确定p53能够滑动——在与双链持续接触的同时沿DNA移动,而不是通过附近碱基之间的一系列跳跃。