scholarly journals 3.1 Å structure of yeast RNA polymerase II elongation complex stalled at a cyclobutane pyrimidine dimer lesion solved using streptavidin affinity grids

2019 ◽  
Vol 207 (3) ◽  
pp. 270-278 ◽  
Author(s):  
Indrajit Lahiri ◽  
Jun Xu ◽  
Bong Gyoon Han ◽  
Juntaek Oh ◽  
Dong Wang ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Pyrimidine Dimer ◽  
Cyclobutane Pyrimidine Dimer ◽  
Elongation Complex

Blockage of RNA polymerase II at a cyclobutane pyrimidine dimer and 6–4 photoproduct

2004 ◽  
Vol 320 (4) ◽  
pp. 1133-1138 ◽  
Author(s):  
Joan Seah Mei Kwei ◽  
Isao Kuraoka ◽  
Katsuyoshi Horibata ◽  
Manabu Ubukata ◽  
Eiry Kobatake ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Pyrimidine Dimer ◽  
Cyclobutane Pyrimidine Dimer

scholarly journals 3.1Å structure of yeast RNA polymerase II elongation complex stalled at a cyclobutane pyrimidine dimer lesion solved using streptavidin affinity grids

2019 ◽  
Author(s):  
Indrajit Lahiri ◽  
Jun Xu ◽  
Bong Gyoon Han ◽  
Juntaek Oh ◽  
Dong Wang ◽  
...  
Keyword(s):  
High Resolution ◽  
Rna Polymerase Ii ◽  
Pyrimidine Dimer ◽  
Water Interface ◽  
Cyclobutane Pyrimidine Dimer ◽  
Elongation Complex ◽  
Pol Ii ◽  
Air Water Interface ◽  
High Resolution Structure ◽  
Resolution Structure

AbstractDespite significant advances in all aspects of single particle cryo-electron microscopy (cryo-EM), specimen preparation still remains a challenge. During sample preparation, macromolecules interact with the air-water interface, which often leads to detrimental effects such as denaturation or adoption of preferred orientations, ultimately hindering structure determination. Randomly biotinylating the protein of interest and then tethering it to a cryo-EM grid coated with two-dimensional crystals of streptavidin (acting as an affinity surface) can prevent the protein from interacting with the air-water interface. Recently, this approach was successfully used to solve a high-resolution structure of a test sample, a bacterial ribosome. However, the general applicability of this method to samples where interaction with the air-water interface is problematic remains to be determined. Here we report a 3.1Å structure of a RNA polymerase II elongation complex stalled at a cyclobutane pyrimidine dimer lesion (Pol II EC(CPD)) solved using streptavidin grids. Our previous attempt to solve this structure using conventional sample preparation methods resulted in a poor quality cryo-EM map due to Pol II EC(CPD)’s adopting a strong preferred orientation. Imaging the same sample on streptavidin grids led to a high-resolution structure with little anisotropy, showing that streptavidin affinity grids could be used as a general strategy to address challenges posed by interaction with the air-water interface.

Allosteric transcription stimulation by RNA polymerase II super elongation complex

2021 ◽  
Author(s):  
Ying Chen ◽  
Seychelle M. Vos ◽  
Christian Dienemann ◽  
Momchil Ninov ◽  
Henning Urlaub ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Elongation Complex ◽  
Super Elongation Complex

scholarly journals Contacts between mammalian RNA polymerase II and the template DNA in a ternary elongation complex

1993 ◽  
Vol 21 (1) ◽  
pp. 113-118 ◽  
Author(s):  
Gretchen A. Rice ◽  
Michael J. Chamberlin ◽  
Caroline M. Kane
Keyword(s):  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Elongation Complex ◽  
Template Dna

Structure of the complete elongation complex of RNA polymerase II with basal factors

Science ◽  
2017 ◽  
Vol 357 (6354) ◽  
pp. 921-924 ◽  
Author(s):  
Haruhiko Ehara ◽  
Takeshi Yokoyama ◽  
Hideki Shigematsu ◽  
Shigeyuki Yokoyama ◽  
Mikako Shirouzu ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Elongation Complex
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