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AFM Analysis of Translocation by EcoR124I
Peter Coxhead1,
Christina Dutta1,
James Smith1,
Olga Kazakova2,
John Van Noort3,
Thijn Van Der Heijden4,
Cees Dekker4 and
Keith Firman1
1 University of
Portsmouth, King Henry Building, King Henry I Street, Portsmouth Hants, PO1
2DY
2 National
Physical Laboratory, Teddington, UK.
3 Leiden University,
The Netherlands.
4 TUDelft, The
Netherlands.
Abstract
Studies
with EcoKI, the archetypical Type IA Restriction-Modification system, have
suggested dimerisation is an integral part of the DNA translocation/cleavage
mechanism.
However single molecule analysis of translocation by the Type IC R-M
system EcoR124I shows no evidence for this effect.
We have also used the AFM to analyse the initiation of translocation.
ATPgS
allows us to ‘trap’ the initial complex and close analysis of images
obtained with the R1-complex
of EcoR124I revealed a small bulge.
We will describe the possible nature of this bulge and its role in the
translocation mechanism.
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