DNA-induced conformational changes in bacteriophage 434 repressor.
Although bacteriophage 434 repressor binds to its specific DNA sites only as a dimer, formation of the dimers in solution occurs at concentrations three orders of magnitude higher than those needed to bind the 434 operator DNA. Our results suggest that both specific and non-specific DNA induce conformational changes in repressor that lead to formation of repressor dimers. The repressor conformational changes induced by DNA occur at concentrations much lower than those needed for binding of repressor, suggesting that the alternative conformations of repressor persist even if the protein is not in direct contact with DNA. Hence, DNA acts in a "catalytic" fashion to induce a steady-state amount of an alternative repressor conformation that has an enhanced affinity for its specific binding site. These findings suggest that the repressor conformer induced by non-specific DNA is the form of the repressor that is optimized for searching for DNA binding sites along non-specific DNA. Upon finding a binding site, the repressor protein undergoes an additional conformational change that allows it to "lock-on" to its specific site.
Ciubotaru
M
M
Department of Biological Sciences, University of New York at Buffalo, 14260-1300, USA.
Bright
F V
FV
Ingersoll
C M
CM
Koudelka
G B
GB
D020106:Acrylamide; D001483:Base Sequence; D001665:Binding Sites; D003090:Coliphages / Q000235:genetics / Q000378:metabolism*; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019281:Dimerization; D019793:Fluorescein; D015345:Oligonucleotide Probes / Q000235:genetics; D009875:Operator Regions, Genetic; D017354:Point Mutation; D011487:Protein Conformation; D020836:Protein Structure, Quaternary; D012097:Repressor Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D014764:Viral Proteins
C063367:434-repressor protein, Bacteriophage 434; D004279:DNA, Viral; D015345:Oligonucleotide Probes; D012097:Repressor Proteins; D014764:Viral Proteins; D020106:Acrylamide; D019793:Fluorescein