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Spt4/5 stimulates transcription elongation through the RNA polymerase clamp coiled-coil motif
Spt5 is the only known RNA polymerase-associated factor that is conserved in all three domains of life. We have solved the structure of the Methanococcus jannaschii Spt4/5 complex by X-ray crystallography, and characterized its function and interaction with the archaeal RNAP in a wholly recombinant...
Autores principales: | , , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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Oxford University Press
2010
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2896526/ https://www.ncbi.nlm.nih.gov/pubmed/20197319 http://dx.doi.org/10.1093/nar/gkq135 |
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author | Hirtreiter, Angela Damsma, Gerke E. Cheung, Alan C. M. Klose, Daniel Grohmann, Dina Vojnic, Erika Martin, Andrew C. R. Cramer, Patrick Werner, Finn |
author_facet | Hirtreiter, Angela Damsma, Gerke E. Cheung, Alan C. M. Klose, Daniel Grohmann, Dina Vojnic, Erika Martin, Andrew C. R. Cramer, Patrick Werner, Finn |
author_sort | Hirtreiter, Angela |
collection | PubMed |
description | Spt5 is the only known RNA polymerase-associated factor that is conserved in all three domains of life. We have solved the structure of the Methanococcus jannaschii Spt4/5 complex by X-ray crystallography, and characterized its function and interaction with the archaeal RNAP in a wholly recombinant in vitro transcription system. Archaeal Spt4 and Spt5 form a stable complex that associates with RNAP independently of the DNA–RNA scaffold of the elongation complex. The association of Spt4/5 with RNAP results in a stimulation of transcription processivity, both in the absence and the presence of the non-template strand. A domain deletion analysis reveals the molecular anatomy of Spt4/5—the Spt5 Nus-G N-terminal (NGN) domain is the effector domain of the complex that both mediates the interaction with RNAP and is essential for its elongation activity. Using a mutagenesis approach, we have identified a hydrophobic pocket on the Spt5 NGN domain as binding site for RNAP, and reciprocally the RNAP clamp coiled-coil motif as binding site for Spt4/5. |
format | Text |
id | pubmed-2896526 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-28965262010-07-06 Spt4/5 stimulates transcription elongation through the RNA polymerase clamp coiled-coil motif Hirtreiter, Angela Damsma, Gerke E. Cheung, Alan C. M. Klose, Daniel Grohmann, Dina Vojnic, Erika Martin, Andrew C. R. Cramer, Patrick Werner, Finn Nucleic Acids Res Nucleic Acid Enzymes Spt5 is the only known RNA polymerase-associated factor that is conserved in all three domains of life. We have solved the structure of the Methanococcus jannaschii Spt4/5 complex by X-ray crystallography, and characterized its function and interaction with the archaeal RNAP in a wholly recombinant in vitro transcription system. Archaeal Spt4 and Spt5 form a stable complex that associates with RNAP independently of the DNA–RNA scaffold of the elongation complex. The association of Spt4/5 with RNAP results in a stimulation of transcription processivity, both in the absence and the presence of the non-template strand. A domain deletion analysis reveals the molecular anatomy of Spt4/5—the Spt5 Nus-G N-terminal (NGN) domain is the effector domain of the complex that both mediates the interaction with RNAP and is essential for its elongation activity. Using a mutagenesis approach, we have identified a hydrophobic pocket on the Spt5 NGN domain as binding site for RNAP, and reciprocally the RNAP clamp coiled-coil motif as binding site for Spt4/5. Oxford University Press 2010-07 2010-03-02 /pmc/articles/PMC2896526/ /pubmed/20197319 http://dx.doi.org/10.1093/nar/gkq135 Text en © The Author(s) 2010. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/2.5 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Nucleic Acid Enzymes Hirtreiter, Angela Damsma, Gerke E. Cheung, Alan C. M. Klose, Daniel Grohmann, Dina Vojnic, Erika Martin, Andrew C. R. Cramer, Patrick Werner, Finn Spt4/5 stimulates transcription elongation through the RNA polymerase clamp coiled-coil motif |
title | Spt4/5 stimulates transcription elongation through the RNA polymerase clamp coiled-coil motif |
title_full | Spt4/5 stimulates transcription elongation through the RNA polymerase clamp coiled-coil motif |
title_fullStr | Spt4/5 stimulates transcription elongation through the RNA polymerase clamp coiled-coil motif |
title_full_unstemmed | Spt4/5 stimulates transcription elongation through the RNA polymerase clamp coiled-coil motif |
title_short | Spt4/5 stimulates transcription elongation through the RNA polymerase clamp coiled-coil motif |
title_sort | spt4/5 stimulates transcription elongation through the rna polymerase clamp coiled-coil motif |
topic | Nucleic Acid Enzymes |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2896526/ https://www.ncbi.nlm.nih.gov/pubmed/20197319 http://dx.doi.org/10.1093/nar/gkq135 |
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