TY - JOUR
T1 - Crystal structure of thermus aquaticus core RNA polymerase at 3.3 å resolution
AU - Zhang, Gongyi
AU - Campbell, Elizabeth A.
AU - Minakhin, Leonid
AU - Richter, Catherine
AU - Severinov, Konstantin
AU - Darst, Seth A.
N1 - Funding Information:
We thank M. Capp and T. Record for the gift of frozen T. aquaticus cell paste; A. Mustaev for stimulating discussions regarding the structure and for experiments to confirm the binding of Pb in the active center metal site; and V. Nikiforov for the extensive alignment of RNAP subunits and for discussions regarding the structure. Some of the DNA and protein sequencing was performed by the Rockefeller University Protein/DNA Technology Center. We are indebted to J. Bonanno and D. Jeruzalmi for assistance with crystallographic calculations; R. Bennet of the Rockefeller University Computing Service for invaluable assistance; J. Kappler and P. Marrack for providing computing facilities; D. Thiel, S. Gruner, and members of the MacCHESS staff, L. Berman and the staff at NSLS X25, and W. Schildkamp and the BioCARS staff at APS for support and assistance in data collection; G. Schneider for the gift of Ta 6 Br 14 ; W. Jahn for the Ir 4 cluster; and J. Goldberg, R. Landick, and S. Nair for helpful discussions. We are indebted to S. K. Burley, J. Kuriyan, R. MacKinnon, and the rest of the Rockefeller University structural biology community for making this project possible. S. A. D. is especially grateful to R. Kornberg, whose own work is a constant inspiration. G. Z. was supported by a National Research Service Award (NIH GM19441-01). E. C. was supported by a Kluge Postdoctoral Fellowship. This work was supported in part by a Burroughs Wellcome Career Development Award to K. S. and a Pew Scholars Award in the Biomedical Sciences to S. A. D.
PY - 1999/9/17
Y1 - 1999/9/17
N2 - The X-ray crystal structure of Thermus aquaticus core RNA polymerase reveals a "crab claw"-shaped molecule with a 27 Å wide internal channel. Located on the back wall of the channel is a Mg2+ ion required for catalytic activity, which is chelated by an absolutely conserved motif from all bacterial and eukaryotic cellular RNA polymerases. The structure places key functional sites, defined by mutational and cross-linking analysis, on the inner walls of the channel in close proximity to the active center Mg2+. Further out from the catalytic center, structural features are found that may be involved in maintaining the melted transcription bubble, clamping onto the RNA product and/or DNA template to assure processivity, and delivering nucleotide substrates to the active center.
AB - The X-ray crystal structure of Thermus aquaticus core RNA polymerase reveals a "crab claw"-shaped molecule with a 27 Å wide internal channel. Located on the back wall of the channel is a Mg2+ ion required for catalytic activity, which is chelated by an absolutely conserved motif from all bacterial and eukaryotic cellular RNA polymerases. The structure places key functional sites, defined by mutational and cross-linking analysis, on the inner walls of the channel in close proximity to the active center Mg2+. Further out from the catalytic center, structural features are found that may be involved in maintaining the melted transcription bubble, clamping onto the RNA product and/or DNA template to assure processivity, and delivering nucleotide substrates to the active center.
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U2 - 10.1016/S0092-8674(00)81515-9
DO - 10.1016/S0092-8674(00)81515-9
M3 - Article
C2 - 10499798
AN - SCOPUS:0033578701
SN - 0092-8674
VL - 98
SP - 811
EP - 824
JO - Cell
JF - Cell
IS - 6
ER -