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, 2006). Because matS sites are found every 35 kb on average, MatP must somehow exert its effects on Ter organization over large distances. To gain insight into the molecular mechanism of MatP-mediated Ter condensation we carried out genetic, biochemical, PF-6463922 solubility dmso and structural studies. These studies unveiled the detailed molecular mechanism by which MatP flexibly links distant matS DNA sites to compact the large Ter MD within enterobacterial chromosomes. A combination of structural, biochemical, genetic, and in?vivo approaches was used to dissect the function of MatP. MatP proteins and matS Ter sites are highly conserved in enterobacteria. Hence, MatP proteins from several Gram-negative bacteria were obtained and purified for crystallographic studies. Structures were obtained of the Yersinia pestis MatP and E.?coli MatP proteins in complex with matS DNA; fluorescence polarization (FP) experiments showed that Y.?pestis MatP binds the matS site with affinity similar to that of E.?coli MatP (Kd?= 2-5?nM) ( Experimental Procedures; Figure?S1). Three different crystal structures of Yersinia pestis MatP-matS were obtained, and one E.?coli MatP-matS structure was solved, providing multiple independent views of the MatP-matS interaction. Two Y.?pestis MatP-matS structures were obtained with the 16-mer matS site, TCGTGACATTGTCACG (where the matS consensus is underlined) to resolutions of 2.80?? and 3.50??, and selleck chemicals llc a structure of Y.?pestis MatP-matS bound to the 23-mer, AGTTCGTGACATTGTCACGAACT was solved to 2.55?? resolution ( Figure?1A; Table 1). Carnitine dehydrogenase A structure of the E.?coli MatP-matS complex was obtained with the 19-mer TTCGTGACATTGTCACGAA to 3.55?? ( Jones et?al., 1991; Br��nger et?al., 1998; Terwilliger and Berendzen, 1999) ( Experimental Procedures; Table 1). The E.?coli and Y.?pestis MatP-matS structures revealed the same overall modular fold comprised of an N-terminal four-helix bundle (Y.?pestis residues 14�C98, E.?coli residues 1�C85) connected to a central �� strand-helix-helix (Y.?pestis residues 104�C145, E.?coli residues 91�C132) and a C-terminal coiled-coil (Y.?pestis residues 146�C160, E.?coli residues 133�C148) ( Figures 1A and 1B). The N-terminal four-helix bundle and strand-helix-helix regions are essentially identical between structures and can be?superimposed with root mean squared deviations (rmsd) for?corresponding C�� atoms of?< 0.8?? for all Y.?pestis and E.?coli structures. By contrast, the coiled-coil region displays flexibility, and its orientation varies between structures. Database searches reveal that MatP contains an overall fold that is?distinct from any previously observed structure; only individual?domains of MatP show similarity to previously solved structures.