Expression, purification and preliminary structural analysis of Escherichia coli MatP in complex with the matS DNA site.
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.
Institut de Biochimie et de Biophysique Moléculaire et Cellulaire, Université de Paris-Sud, UMR8619 du CNRS, Bâtiment 430, 91405 Orsay, France.
Dominique
D
Durand
Ines
I
Li de la Sierra-Gallay
Mark A
MA
Brooks
Andrew W
AW
Thompson
Noureddine
N
Lazar
Johnny
J
Lisboa
Herman
H
van Tilbeurgh
Sophie
S
Quevillon-Cheruel
D000595:Amino Acid Sequence; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D004269:DNA, Bacterial / Q000737:chemistry* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D004926:Escherichia coli / Q000737:chemistry* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D015870:Gene Expression; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment
D002868:Chromosomal Proteins, Non-Histone; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C533037:MatP protein, E coli