Purification and characterization of SopA and SopB proteins essential for F plasmid partitioning.
Mini-F plasmid has the trans-acting genes sopA and sopB and the cis-acting site sopC which are essential for accurate partitioning of plasmid DNA molecules into both daughter cells. In this study, we purified independently SopA and SopB proteins, analyzed the in vitro DNA-binding activity of these proteins by the gel retardation assay, and determined the precise binding sites of DNA by the footprinting method. SopA binds to four repeated sequences (CTTTGC) located in the promoter-operator region of the sopAB operon. The SopA binding activity is enhanced by the addition of SopB protein. SopB protein itself does not bind to this DNA region. These results suggest that the complex of SopA and SopB proteins autoregulate the expression of the sopA-sopB operon. On the other hand, SopB protein binds to the sopC region, in which 12 direct repeats of 43-base pairs nucleotides exist. SopB protein recognizes the inverted repeats of 7 base pairs in each direct repeats. SopA protein does not affect the SopB binding activity to the sopC DNA segment.
Mori
H
H
Department of Molecular Genetics, Kumamoto University Medical School, Japan.
Mori
Y
Y
Ichinose
C
C
Niki
H
H
Ogura
T
T
Kato
A
A
Hiraga
S
S
D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D001483:Base Sequence; D003850:Deoxyribonuclease I; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D005796:Genes; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D009710:Nucleotide Mapping; D009876:Operon; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C060972:SopB protein, Bacteria; C489223:SopB protein, E coli; D003850:Deoxyribonuclease I; C060971:SopA protein, Bacteria