scholarly journals Detection of Reaction Intermediates in Mg 2+ -Dependent DNA Synthesis and RNA Degradation by Time-Resolved X-Ray Crystallography

Author(s):  
Nadine L. Samara ◽  
Yang Gao ◽  
Jinjun Wu ◽  
Wei Yang
2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Nicholas Chim ◽  
Roman A. Meza ◽  
Anh M. Trinh ◽  
Kefan Yang ◽  
John C. Chaput

AbstractThe mechanism of DNA synthesis has been inferred from static structures, but the absence of temporal information raises longstanding questions about the order of events in one of life’s most central processes. Here we follow the reaction pathway of a replicative DNA polymerase using time-resolved X-ray crystallography to elucidate the order and transition between intermediates. In contrast to the canonical model, the structural changes observed in the time-lapsed images reveal a catalytic cycle in which translocation precedes catalysis. The translocation step appears to follow a push-pull mechanism where the O-O1 loop of the finger subdomain acts as a pawl to facilitate unidirectional movement along the template with conserved tyrosine residues 714 and 719 functioning as tandem gatekeepers of DNA synthesis. The structures capture the precise order of critical events that may be a general feature of enzymatic catalysis among replicative DNA polymerases.


1997 ◽  
Vol 30 (5) ◽  
pp. 555-556 ◽  
Author(s):  
G. Kurisu ◽  
A. Sugimoto ◽  
Y. Kai ◽  
S. Harada

Crystals ◽  
2020 ◽  
Vol 10 (10) ◽  
pp. 854
Author(s):  
Ki Hyun Nam

Radiation damage and cryogenic sample environment are an experimental limitation observed in the traditional X-ray crystallography technique. However, the serial crystallography (SX) technique not only helps to determine structures at room temperature with minimal radiation damage, but it is also a useful tool for profound understanding of macromolecules. Moreover, it is a new tool for time-resolved studies. Over the past 10 years, various sample delivery techniques and data collection strategies have been developed in the SX field. It also has a wide range of applications in instruments ranging from the X-ray free electron laser (XFEL) facility to synchrotrons. The importance of the various approaches in terms of the experimental techniques and a brief review of the research carried out in the field of SX has been highlighted in this editorial.


2006 ◽  
Vol 153 (3) ◽  
pp. 300-306 ◽  
Author(s):  
William R. Wikoff ◽  
James F. Conway ◽  
Jinghua Tang ◽  
Kelly K. Lee ◽  
Lu Gan ◽  
...  

2004 ◽  
Vol 147 (3) ◽  
pp. 235-246 ◽  
Author(s):  
Friedrich Schotte ◽  
Jayashree Soman ◽  
John S. Olson ◽  
Michael Wulff ◽  
Philip A. Anfinrud

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