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T7 RNA polymerase functions in vitro without clustering.

Abstract:
Many nucleic acid polymerases function in clusters known as factories. We investigate whether the RNA polymerase (RNAP) of phage T7 also clusters when active. Using 'pulldowns' and fluorescence correlation spectroscopy we find that elongation complexes do not interact in vitro with a K(d)<1 micrometre. Chromosome conformation capture also reveals that genes located 100 kb apart on the E. coli chromosome do not associate more frequently when transcribed by T7 RNAP. We conclude that if clustering does occur in vivo, it must be driven by weak interactions, or mediated by a phage-encoded protein.
Publication status:
Published
Peer review status:
Peer reviewed

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Publisher copy:
10.1371/journal.pone.0040207

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Institution:
University of Oxford
Role:
Author
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Institution:
University of Oxford
Role:
Author
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Institution:
University of Oxford
Division:
MPLS
Department:
Physics
Sub department:
Condensed Matter Physics
Role:
Author
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Institution:
University of Oxford
Division:
MSD
Department:
Pathology Dunn School
Role:
Author


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Funding agency for:
Kapanidis, A
Grant:
BB/H01795X/1
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Funding agency for:
Kapanidis, A
Grant:
BB/H01795X/1
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Funding agency for:
Finan, K
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Funding agency for:
Finan, K
Torella, J
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Funding agency for:
Finan, K
Cook, P


Publisher:
Public Library of Science
Journal:
PloS one More from this journal
Volume:
7
Issue:
7
Article number:
e40207
Publication date:
2012-01-01
DOI:
EISSN:
1932-6203
ISSN:
1932-6203


Language:
English
Keywords:
UUID:
uuid:11113cc0-25b9-48c1-8713-5d4d12133986
Local pid:
pubs:342278
Source identifiers:
342278
Deposit date:
2012-12-19

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