Abstract
RNA polymerase functions like a molecular motor that can convert chemical energy into the work of strand separation and translocation along the DNA during transcription. The structures of phage T7 RNA polymerase in an elongation phase substrate complex that includes the incoming nucleoside triphosphate and a pretranslocation product complex that includes the product pyrophosphate (PPi) are described here. These structures and the previously determined posttranslocation elongation complex demonstrate that two enzyme conformations exist during a cycle of single nucleotide addition. One orientation of a five-helix subdomain is stabilized by the phosphates of either the incoming NTP or by the product PPi. A second orientation of this subdomain is stable in their absence and is associated with translocation of the heteroduplex product as well as strand separation of the downstream DNA. We propose that the dissociation of the product PPi after nucleotide addition produces the protein conformational change resulting in translocation and strand separation.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 393-404 |
| Number of pages | 12 |
| Journal | Cell |
| Volume | 116 |
| Issue number | 3 |
| DOIs | |
| State | Published - Feb 6 2004 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Cell Biology
- Molecular Biology
Cite this
The structural mechanism of translocation and helicase activity in T7 RNA polymerase. / Yin, Yuhui; Steitz, Thomas A.
In: Cell, Vol. 116, No. 3, 06.02.2004, p. 393-404.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - The structural mechanism of translocation and helicase activity in T7 RNA polymerase
AU - Yin, Yuhui
AU - Steitz, Thomas A.
PY - 2004/2/6
Y1 - 2004/2/6
N2 - RNA polymerase functions like a molecular motor that can convert chemical energy into the work of strand separation and translocation along the DNA during transcription. The structures of phage T7 RNA polymerase in an elongation phase substrate complex that includes the incoming nucleoside triphosphate and a pretranslocation product complex that includes the product pyrophosphate (PPi) are described here. These structures and the previously determined posttranslocation elongation complex demonstrate that two enzyme conformations exist during a cycle of single nucleotide addition. One orientation of a five-helix subdomain is stabilized by the phosphates of either the incoming NTP or by the product PPi. A second orientation of this subdomain is stable in their absence and is associated with translocation of the heteroduplex product as well as strand separation of the downstream DNA. We propose that the dissociation of the product PPi after nucleotide addition produces the protein conformational change resulting in translocation and strand separation.
AB - RNA polymerase functions like a molecular motor that can convert chemical energy into the work of strand separation and translocation along the DNA during transcription. The structures of phage T7 RNA polymerase in an elongation phase substrate complex that includes the incoming nucleoside triphosphate and a pretranslocation product complex that includes the product pyrophosphate (PPi) are described here. These structures and the previously determined posttranslocation elongation complex demonstrate that two enzyme conformations exist during a cycle of single nucleotide addition. One orientation of a five-helix subdomain is stabilized by the phosphates of either the incoming NTP or by the product PPi. A second orientation of this subdomain is stable in their absence and is associated with translocation of the heteroduplex product as well as strand separation of the downstream DNA. We propose that the dissociation of the product PPi after nucleotide addition produces the protein conformational change resulting in translocation and strand separation.
UR - http://www.scopus.com/inward/record.url?scp=1342313235&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=1342313235&partnerID=8YFLogxK
U2 - 10.1016/S0092-8674(04)00120-5
DO - 10.1016/S0092-8674(04)00120-5
M3 - Article
C2 - 15016374
AN - SCOPUS:1342313235
VL - 116
SP - 393
EP - 404
JO - Cell
JF - Cell
SN - 0092-8674
IS - 3
ER -