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Similar smFRET histograms and periodic unwinding time traces were obtained, implying that the periodic G4 unwinding by a Pif1 monomer may be general and not specific to sub-species of G4 structures.
(B ) Representative unwinding time course for labeled dsRNA (125 nM) by the helicase core of Mss116 (2 μM) measured at 5 mM ATP-Mg2+.
A 10-fold difference in the unwinding time from 0% GC sequence to 80% GC sequence was observed (1 and 10 s, respectively; Figure 1C).
The translocation rate could be obtained by plotting the unwinding time as a function of length: the mutant gives a slightly slower rate (19 bp/s) than the wild type (30 bp/s).
(C ) Representative unwinding time course for labeled dsRNA (125 nM) by the helicase core of Mss116 (2 μM) measured at 5 mM CTP-Mg2+ with samples resolved as in (B ).
To quantify the unwinding behavior, we measured the total unwinding time, which is the duration of the time interval from the moment FRET starts to decrease until the moment the total fluorescence signal disappears, and plotted the distributions.
Similar(49)
Under the monomer condition, unwinding curves for RD31 can be fit to a single exponential function, yielding the unwinding times tunwind = 6.3 ± 0.9 and 1.6 ± 0.2 min at 20 μM and 1 mM ATP, respectively.
Data were fit to a single exponential equation (fraction unwound = A{1-exp(- ku t)}); A, amplitude of the reaction; ku, apparent first-order rate constant for unwinding; t, time using GraphPad Prism (GraphPad Software , Inc.
These average unwinding rates include time spent in unwinding the 40 bp fork DNA and time spent in any paused states.
According to The Sleep Council – a non-profit organisation that aims to help people improve their sleeping habits – it's important to spend time unwinding before going to bed.
"We're going to Center Parcs for a few days in the run-up to Christmas, to see Santa and spend time unwinding as a family, instead of shooting around the shops.
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CEO of Professional Science Editing for Scientists @ prosciediting.com