Gene machines

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In vivo single-molecule biochemistry of chromosome replication and segregation in bacteria.

MOLECULAR BIOLOGY OF THE CELL 23 (2012)

Authors:

D Sherratt, R Reyes-Lamothe, A Badrinaryanan, E Nicolas, O Henry, A Kapanidis, S Uphoff, M Leake
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Measuring enzyme copy numbers and single-molecule reaction rates in living cells.

MOLECULAR BIOLOGY OF THE CELL 23 (2012)

Authors:

S Uphoff, R Reyes-Lamothe, F Garza de Leon, M Leake, D Sherratt, A Kapanidis
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Single-molecule FRET measurements in bacterial cells

FEBS JOURNAL 279 (2012) 513-513

Authors:

L Aigrain, R Crawford, J Torella, A Plochowietz, A Kapanidis
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Super-Resolution Fluorescence Microscopy of Transcription Sites in E. Coli

BIOPHYSICAL JOURNAL 102:3 (2012) 229A-229A

Authors:

Kieran R Finan, Mike Heilemann, Ulrike Endesfelder, Seamus Holden, Peter R Cook, Achilles Kapanidis
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T7 RNA polymerase functions in vitro without clustering.

PLoS One 7:7 (2012) e40207

Authors:

Kieran Finan, Joseph P Torella, Achillefs N Kapanidis, Peter R Cook

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 µM. 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.
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