Cryo-EM structure of MukBEF reveals DNA loop entrapment at chromosomal unloading sites

Abstract

The ring-like structural maintenance of chromosomes (SMC) complex MukBEF folds the genome of Escherichia coli and related bacteria into large loops, presumably by active DNA loop extrusion. MukBEF activity within the replication terminus macrodomain is suppressed by the sequence specific unloader MatP. Here we present the complete atomic structure of MukBEF in complex with MatP and DNA as determined by electron cryomicroscopy (cryo-EM). The complex binds two distinct DNA double helices corresponding to the arms of a plectonemic loop. MatP-bound DNA threads through the MukBEF ring, while the second DNA is clamped by the kleisin MukF, MukE and the MukB ATPase heads. Combinatorial cysteine cross-linking confirms this topology of DNA loop entrapment in vivo. Our findings illuminate how a class of near-ubiquitous DNA organizers with important roles in genome maintenance interacts with the bacterial chromosome. HighlightsO_LIComplete atomic structures of the bacterial SMC complex MukBEF on and off DNA. C_LIO_LIMukBEF entraps two DNA double helices when bound to the unloader MatP. C_LIO_LIIn vivo topology of DNA loop entrapment determined by cysteine cross-linking. C_LIO_LIArms of the DNA loop thread through separate compartments of MukBEF. C_LI Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=189 SRC="FIGDIR/small/450292v1_ufig1.gif" ALT="Figure 1"> View larger version (44K): org.highwire.dtl.DTLVardef@2fdadorg.highwire.dtl.DTLVardef@5c8cc3org.highwire.dtl.DTLVardef@1d8ecforg.highwire.dtl.DTLVardef@8af8b8_HPS_FORMAT_FIGEXP M_FIG C_FIG