TY - JOUR
T1 - Structural basis of transcription
T2 - Nucleotide selection by rotation in the RNA polymerase II active center
AU - Westover, Kenneth D.
AU - Bushnell, David A.
AU - Kornberg, Roger D.
N1 - Funding Information:
K.D.W. was supported by the Medical Scientist Training Program. This research was supported by NIH grants GM49985 and AI21144 to R.D.K.
PY - 2004/11/12
Y1 - 2004/11/12
N2 - Binding of a ribonucleoside triphosphate to an RNA polymerase II transcribing complex, with base pairing to the template DNA, was revealed by X-ray crystallography. Binding of a mismatched nucleoside triphosphate was also detected, but in an adjacent site, inverted with respect to the correctly paired nucleotide. The results are consistent with a two-step mechanism of nucleotide selection, with initial binding to an entry (E) site beneath the active center in an inverted orientation, followed by rotation into the nucleotide addition (A) site for pairing with the template DNA. This mechanism is unrelated to that of single subunit RNA polymerases and so defines a new paradigm for the large, multisubunit enzymes. Additional findings from these studies include a third nucleotide binding site that may define the length of backtracked RNA; DNA double helix unwinding in advance of the polymerase active center; and extension of the diffraction limit of RNA polymerase II crystals to 2.3 Å.
AB - Binding of a ribonucleoside triphosphate to an RNA polymerase II transcribing complex, with base pairing to the template DNA, was revealed by X-ray crystallography. Binding of a mismatched nucleoside triphosphate was also detected, but in an adjacent site, inverted with respect to the correctly paired nucleotide. The results are consistent with a two-step mechanism of nucleotide selection, with initial binding to an entry (E) site beneath the active center in an inverted orientation, followed by rotation into the nucleotide addition (A) site for pairing with the template DNA. This mechanism is unrelated to that of single subunit RNA polymerases and so defines a new paradigm for the large, multisubunit enzymes. Additional findings from these studies include a third nucleotide binding site that may define the length of backtracked RNA; DNA double helix unwinding in advance of the polymerase active center; and extension of the diffraction limit of RNA polymerase II crystals to 2.3 Å.
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U2 - 10.1016/j.cell.2004.10.016
DO - 10.1016/j.cell.2004.10.016
M3 - Article
C2 - 15537538
AN - SCOPUS:8344234112
SN - 0092-8674
VL - 119
SP - 481
EP - 489
JO - Cell
JF - Cell
IS - 4
ER -