Fast, DNA-sequence independent translocation by FtsK in a single-molecule experiment.
Escherichia coli FtsK is an essential cell division protein, which is thought to pump chromosomal DNA through the closing septum in an oriented manner by following DNA sequence polarity. Here, we perform single-molecule measurements of translocation by FtsK50C, a derivative that functions as a DNA translocase in vitro. FtsK50C translocation follows Michaelis-Menten kinetics, with a maximum speed of approximately 6.7 kbp/s. We present results on the effect of applied force on the speed, distance translocated, and the mean times during and between protein activity. Surprisingly, we observe that FtsK50C can spontaneously reverse its translocation direction on a fragment of E. coli chromosomal DNA, indicating that DNA sequence is not the sole determinant of translocation direction. We conclude that in vivo polarization of FtsK translocation could require the presence of cofactors; alternatively, we propose a model in which tension in the DNA directs FtsK translocation.
Saleh
Omar A
OA
Laboratoire de Physique Statistique et Département de Biologie, Ecole Normale Supérieure, Paris, France.
Pérals
Corine
C
Barre
François-Xavier
FX
Allemand
Jean-François
JF
D004247:DNA / Q000235:genetics*; D029968:Escherichia coli Proteins; D008565:Membrane Proteins / Q000502:physiology*; D021381:Protein Transport
D029968:Escherichia coli Proteins; C096452:FtsK protein, E coli; D008565:Membrane Proteins; D004247:DNA