Molecular architecture and conformational flexibility of human RNA polymerase II.
Transcription by RNA polymerase II (RNAPII) is a central process in eukaryotic gene regulation. While atomic details exist for the yeast RNAPII, characterization of the human complex lags behind, mostly due to the inability to obtain large quantities of purified material. Although the complexes have the same protein composition and high sequence similarity, understanding of transcription and of transcription-coupled DNA repair (TCR) in humans will require the use of human proteins in structural studies. We have used cryo-electron microscopy, image reconstruction, and variance analysis to characterize the structure and dynamics of human RNAPII (hRNAPII). Our studies show that hRNAPII in solution parallels the conformational flexibility of the yeast structures crystallized in different states but also illustrate a more extensive conformational range with potential biological significance. This hRNAPII study will serve as a structural platform to build up higher-order transcription and TCR complexes and to gain information that may be unique to the human RNAPII system.
Kostek
Seth A
SA
Molecular and Cell Biology Department, University of California, Berkeley, Berkeley, California 94720, USA.
Grob
Patricia
P
De Carlo
Sacha
S
Lipscomb
J Slaton
JS
Garczarek
Florian
F
Nogales
Eva
E
D020285:Cryoelectron Microscopy; D005656:Fungal Proteins / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D021621:Imaging, Three-Dimensional; D008854:Microscopy, Electron; D008958:Models, Molecular; D008968:Molecular Conformation; D011487:Protein Conformation; D012319:RNA Polymerase II / Q000737:chemistry*; D014158:Transcription, Genetic
D005656:Fungal Proteins; D012319:RNA Polymerase II