E. coli DnaB bound to two DnaG C-terminal domains, ssDNA, ADP and AlF4

Sequence of conformation changes of DnaB helicase during DNA unwinding and priming in Escherichia coli

20 mM Tris-HCl, pH 7.6, 100 mM NaCl, 5 mM MgCl2, 3 mM DTT, 0.25 mM EDTA and 100 micromolar ADP.

E. coli DnaB DnaG C-terminal domain bound to ssDNA

Grids covered with a 2 nm continuous carbon film. Glow-discharged grids were floated on 40 microL drops of a 0.1% (w/v) aqueous solution of poly-lysine hydrobromide (MW 30-70 kDa; Polysciences, Inc.) for two min, briefly rinsed with MilliQ water 4 times and then allow to air dry.

3 microL of sample was applied to glow-discharged grids. Grids were blotted at 6 degrees C for 3.5 s with no extra blot force.

DNA (5'-D(*TP*TP*TP*TP*TP*TP*TP*TP*TP*TP*TP*TP*TP*TP*TP*TP*TP*TP*TP*T)-3')

E. COLI DNAB BOUND TO DNAG C-TERMINAL DOMAIN AND SSDNA

helicase, SF4, AAA+, REPLICATION-DNA complex