Details
| Original language | English |
|---|---|
| Pages (from-to) | 863-866 |
| Number of pages | 4 |
| Journal | SCIENCE |
| Volume | 356 |
| Issue number | 6340 |
| Publication status | Published - 26 May 2017 |
| Externally published | Yes |
Abstract
All cellular RNA polymerases (RNAPs), from those of bacteria to those of man, possess a clamp that can open and close, and it has been assumed that the open RNAP separates promoter DNA strands and then closes to establish a tight grip on the DNA template. Here, we resolve successive motions of the initiating bacterial RNAP by studying real-time signatures of fluorescent reporters placed on RNAP and DNA in the presence of ligands locking the clamp in distinct conformations. We report evidence for an unexpected and obligatory step early in the initiation involving a transient clamp closure as a prerequisite for DNA melting. We also present a 2.6-angstrom crystal structure of a late-initiation intermediate harboring a rotationally unconstrained downstream DNA duplex within the open RNAP active site cleft. Our findings explain how RNAP thermal motions control the promoter search and drive DNA melting in the absence of external energy sources.
ASJC Scopus subject areas
Cite this
- Standard
- Harvard
- Apa
- Vancouver
- BibTeX
- RIS
In: SCIENCE, Vol. 356, No. 6340, 26.05.2017, p. 863-866.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - RNA polymerase motions during promoter melting
AU - Feklistov, Andrey
AU - Bae, Brian
AU - Hauver, Jesse
AU - Lass-Napiorkowska, Agnieszka
AU - Kalesse, Markus
AU - Glaus, Florian
AU - Altmann, Karl Heinz
AU - Heyduk, Tomasz
AU - Landick, Robert
AU - Darst, Seth A.
N1 - Publisher Copyright: Copyright 2016 by the American Association for the Advancement of Science; all rights reserved. Copyright: Copyright 2017 Elsevier B.V., All rights reserved.
PY - 2017/5/26
Y1 - 2017/5/26
N2 - All cellular RNA polymerases (RNAPs), from those of bacteria to those of man, possess a clamp that can open and close, and it has been assumed that the open RNAP separates promoter DNA strands and then closes to establish a tight grip on the DNA template. Here, we resolve successive motions of the initiating bacterial RNAP by studying real-time signatures of fluorescent reporters placed on RNAP and DNA in the presence of ligands locking the clamp in distinct conformations. We report evidence for an unexpected and obligatory step early in the initiation involving a transient clamp closure as a prerequisite for DNA melting. We also present a 2.6-angstrom crystal structure of a late-initiation intermediate harboring a rotationally unconstrained downstream DNA duplex within the open RNAP active site cleft. Our findings explain how RNAP thermal motions control the promoter search and drive DNA melting in the absence of external energy sources.
AB - All cellular RNA polymerases (RNAPs), from those of bacteria to those of man, possess a clamp that can open and close, and it has been assumed that the open RNAP separates promoter DNA strands and then closes to establish a tight grip on the DNA template. Here, we resolve successive motions of the initiating bacterial RNAP by studying real-time signatures of fluorescent reporters placed on RNAP and DNA in the presence of ligands locking the clamp in distinct conformations. We report evidence for an unexpected and obligatory step early in the initiation involving a transient clamp closure as a prerequisite for DNA melting. We also present a 2.6-angstrom crystal structure of a late-initiation intermediate harboring a rotationally unconstrained downstream DNA duplex within the open RNAP active site cleft. Our findings explain how RNAP thermal motions control the promoter search and drive DNA melting in the absence of external energy sources.
UR - http://www.scopus.com/inward/record.url?scp=85019707365&partnerID=8YFLogxK
U2 - 10.1126/science.aam7858
DO - 10.1126/science.aam7858
M3 - Article
C2 - 28546214
AN - SCOPUS:85019707365
VL - 356
SP - 863
EP - 866
JO - SCIENCE
JF - SCIENCE
SN - 0036-8075
IS - 6340
ER -