Expression, purification and preliminary structural analysis of Escherichia coli MatP in complex with the matS DNA site

Abstract

The Escherichia coli chromosome is organized into four macrodomains which are found in the replication-origin region (Ori), at the terminus (Ter) and on both its sides (Right and Left). The localization of the macrodomains is subject to programmed changes during the cell cycle. The compaction of the 800 kb Ter macrodomain relies on the binding of the MatP protein to a 13 bp matS motif repeated 23 times. MatP is a small DNA-binding protein of about 18 kDa that shares homology in its C-terminal region with the ribbon-helix-helix (RHH) motifs present in regulatory DNA-binding proteins such as CopG. In order to understand the DNA-compaction mechanism of MatP at an atomic level, it was decided to study the structure of apo MatP and of the nucleoprotein complex MatP-matS by both X-ray diffraction and SAXS analysis. It was demonstrated that MatP forms dimers that bind a single matS motif. Complete native X-ray data sets were collected and phasing of the diffraction data is under way. © 2012 International Union of Crystallography All rights reserved.