Structural basis for the transition from initiation to elongation transcription in T7 RNA polymerase

Yuhui Yin, Thomas A. Steitz

Research output: Contribution to journalArticle

241 Citations (Scopus)

Abstract

To make messenger RNA transcripts, bacteriophage T7 RNA polymerase (T7 RNAP) undergoes a transition from an initiation phase, which only makes short RNA fragments, to a stable elongation phase. We have determined at 2.1 angstrom resolution the crystal structure of a T7 RNAP elongation complex with 30 base pairs of duplex DNA containing a "transcription bubble" interacting with a 17-nucleotide RNA transcript. The transition from an initiation to an elongation complex is accompanied by a major refolding of the amino-terminal 300 residues. This results in loss of the promoter binding site, facilitating promoter clearance, and creates a tunnel that surrounds the RNA transcript after it peels off a seven-base pair heteroduplex. Formation of the exit tunnel explains the enhanced processivity of the elongation complex. Downstream duplex DNA binds to the fingers domain, and its orientation relative to upstream DNA in the initiation complex implies an unwinding that could facilitate formation of the open promoter complex.

Original languageEnglish (US)
Pages (from-to)1387-1395
Number of pages9
JournalScience
Volume298
Issue number5597
DOIs
StatePublished - Nov 15 2002
Externally publishedYes

Fingerprint

RNA
Base Pairing
DNA
RNA Phages
Phase Transition
Fingers
Nucleotides
Binding Sites
Messenger RNA
bacteriophage T7 RNA polymerase

ASJC Scopus subject areas

  • General

Cite this

Structural basis for the transition from initiation to elongation transcription in T7 RNA polymerase. / Yin, Yuhui; Steitz, Thomas A.

In: Science, Vol. 298, No. 5597, 15.11.2002, p. 1387-1395.

Research output: Contribution to journalArticle

@article{ab9b1a16c21243269d1ad985774482cd,
title = "Structural basis for the transition from initiation to elongation transcription in T7 RNA polymerase",
abstract = "To make messenger RNA transcripts, bacteriophage T7 RNA polymerase (T7 RNAP) undergoes a transition from an initiation phase, which only makes short RNA fragments, to a stable elongation phase. We have determined at 2.1 angstrom resolution the crystal structure of a T7 RNAP elongation complex with 30 base pairs of duplex DNA containing a {"}transcription bubble{"} interacting with a 17-nucleotide RNA transcript. The transition from an initiation to an elongation complex is accompanied by a major refolding of the amino-terminal 300 residues. This results in loss of the promoter binding site, facilitating promoter clearance, and creates a tunnel that surrounds the RNA transcript after it peels off a seven-base pair heteroduplex. Formation of the exit tunnel explains the enhanced processivity of the elongation complex. Downstream duplex DNA binds to the fingers domain, and its orientation relative to upstream DNA in the initiation complex implies an unwinding that could facilitate formation of the open promoter complex.",
author = "Yuhui Yin and Steitz, {Thomas A.}",
year = "2002",
month = "11",
day = "15",
doi = "10.1126/science.1077464",
language = "English (US)",
volume = "298",
pages = "1387--1395",
journal = "Science",
issn = "0036-8075",
publisher = "American Association for the Advancement of Science",
number = "5597",

}

TY - JOUR

T1 - Structural basis for the transition from initiation to elongation transcription in T7 RNA polymerase

AU - Yin, Yuhui

AU - Steitz, Thomas A.

PY - 2002/11/15

Y1 - 2002/11/15

N2 - To make messenger RNA transcripts, bacteriophage T7 RNA polymerase (T7 RNAP) undergoes a transition from an initiation phase, which only makes short RNA fragments, to a stable elongation phase. We have determined at 2.1 angstrom resolution the crystal structure of a T7 RNAP elongation complex with 30 base pairs of duplex DNA containing a "transcription bubble" interacting with a 17-nucleotide RNA transcript. The transition from an initiation to an elongation complex is accompanied by a major refolding of the amino-terminal 300 residues. This results in loss of the promoter binding site, facilitating promoter clearance, and creates a tunnel that surrounds the RNA transcript after it peels off a seven-base pair heteroduplex. Formation of the exit tunnel explains the enhanced processivity of the elongation complex. Downstream duplex DNA binds to the fingers domain, and its orientation relative to upstream DNA in the initiation complex implies an unwinding that could facilitate formation of the open promoter complex.

AB - To make messenger RNA transcripts, bacteriophage T7 RNA polymerase (T7 RNAP) undergoes a transition from an initiation phase, which only makes short RNA fragments, to a stable elongation phase. We have determined at 2.1 angstrom resolution the crystal structure of a T7 RNAP elongation complex with 30 base pairs of duplex DNA containing a "transcription bubble" interacting with a 17-nucleotide RNA transcript. The transition from an initiation to an elongation complex is accompanied by a major refolding of the amino-terminal 300 residues. This results in loss of the promoter binding site, facilitating promoter clearance, and creates a tunnel that surrounds the RNA transcript after it peels off a seven-base pair heteroduplex. Formation of the exit tunnel explains the enhanced processivity of the elongation complex. Downstream duplex DNA binds to the fingers domain, and its orientation relative to upstream DNA in the initiation complex implies an unwinding that could facilitate formation of the open promoter complex.

UR - http://www.scopus.com/inward/record.url?scp=0037112082&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037112082&partnerID=8YFLogxK

U2 - 10.1126/science.1077464

DO - 10.1126/science.1077464

M3 - Article

VL - 298

SP - 1387

EP - 1395

JO - Science

JF - Science

SN - 0036-8075

IS - 5597

ER -