Crystallization and preliminary X-ray crystallographic analyses ofThermus thermophilusbacktracked RNA polymerase

Yuko Murayama, Shun-ichi Sekine, Shigeyuki Yokoyama
2013 Acta Crystallographica. Section F : Structural Biology and Crystallization Communications  
DNA-dependent RNA polymerase (RNAP) synthesizes RNA complementary to the template DNA. During transcript elongation, RNAP often undergoes backward translocation ('backtracking') by dissociating the 3 0 end of the nascent RNA transcript from the template DNA. While the backtracked state of RNAP is inactive in RNA elongation, it actively hydrolyses the RNA 3 0 end to regenerate the active elongation complex. To study the structural basis of the backtracked state and its cleavage activity, two
more » ... tracked RNAP complexes were reconstituted by assembling Thermus thermophilus RNAP with designed nucleic acid scaffolds. The reconstituted backtracked complexes were active in the transcript-cleavage reaction. These complexes were crystallized and X-ray diffraction data sets were obtained at resolutions of 3.4 and 3.7 Å .
doi:10.1107/s1744309113000055 pmid:23385762 pmcid:PMC3564623 fatcat:xmzl2f5xvrf2zjinec2wsoehlq