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Published September 17, 1999 | public
Journal Article

Yeast RNA Polymerase II at 5 Å Resolution

Abstract

Appropriate treatment of X-ray diffraction from an unoriented 18-heavy atom cluster derivative of a yeast RNA polymerase II crystal gave significant phase information to 5 Å resolution. The validity of the phases was shown by close similarity of a 6 Å electron density map to a 16 Å molecular envelope of the polymerase from electron crystallography. Comparison of the 6 Å X-ray map with results of electron crystallography of a paused transcription elongation complex suggests functional roles for two mobile protein domains: the tip of a flexible arm forms a downstream DNA clamp; and a hinged domain may serve as an RNA clamp, enclosing the transcript from about 8–18 residues upstream of the 3′-end in a tunnel.

Additional Information

© 1999 Cell Press. Under an Elsevier user license. Received 7 July 1999, Revised 10 August 1999. We thank Michael Pope (George Washington University) for tungsten clusters; Henry Bellemy of SSRL for advice on anomalous scattering experiments; Gunter Schneider (Karolinska Institute, Sweden) for the Ta_6Br_(14) complex; Robert Huber (Max-Planck-Institute, Germany) for various heavy atom compounds; Ken Ng, Anand Kolatkar, and Joseph Wedekind for assistance with X-ray facilities; Mincheng Zhang for help with protein purification; and Katherine Nolan for help in the preparation of monoclonal antibody. J. F. acknowledges fellowship support from the Universitywide AIDS Research Program, University of California (#F96-ST-015) and American Cancer Society (#PF-99-112-01-GMC). The contribution of A. L. G. was sponsored by USAMRC Breast Cancer Initiative, DAMD17-97-1-7099 and does not necessarily reflect the position or policy of the government. Work done at SSRL is supported by the NIH (National Center for Research Resources, Biomedical Technology Program) and by the Department of Energy. This research was supported by NIH grant GM49985 to R. D. K.

Additional details

Created:
August 19, 2023
Modified:
October 19, 2023