E.coli MukB protein involved in chromosome partition forms a homodimer with a rod-and-hinge structure having DNA binding and ATP/GTP binding activities.
mukB mutants of Escherichia coli are defective in the correct partitioning of replicated chromosomes. This results in the appearance of normal-sized anucleate (chromosome-less) cells during cell proliferation. Based on the nucleotide sequence of the mukB gene, the MukB protein of 177 kDa was predicted to be a filamentous protein with globular domains at the ends, and also having DNA binding and nucleotide binding abilities. Here we present evidence that the purified MukB protein possesses these characteristics. MukB forms a homodimer with a rod-and-hinge structure having a pair of large, C-terminal globular domains at one end and a pair of small, N-terminal globular domains at the opposite end; it tends to bend at a middle hinge site of the rod section. Chromatography in a DNA-cellulose column and the gel retardation assay revealed that MukB possesses DNA binding activity. Photoaffinity cross-linking experiments showed that MukB binds to ATP and GTP in the presence of Zn2+. Throughout the purification steps, acyl carrier protein was co-purified with MukB.
Niki
H
H
Department of Molecular Cell Biology, Kumamoto University School of Medicine, Japan.
Imamura
R
R
Kitaoka
M
M
Yamanaka
K
K
Ogura
T
T
Hiraga
S
S
D000213:Acyl Carrier Protein / Q000302:isolation & purification; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism / Q000648:ultrastructure; D015153:Blotting, Western; D002850:Chromatography, Gel; D002868:Chromosomal Proteins, Non-Histone; D002876:Chromosomes, Bacterial; D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism / Q000648:ultrastructure; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins; D006160:Guanosine Triphosphate / Q000378:metabolism*; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D011487:Protein Conformation
D000213:Acyl Carrier Protein; D001426:Bacterial Proteins; D002868:Chromosomal Proteins, Non-Histone; D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C067035:MukB protein, E coli; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate