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Mutational and physical analysis of F plasmid traY protein binding to oriT
Author(s) -
Luo Yanan,
Gao Qian,
Deonier Richard C.
Publication year - 1994
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1994.tb00327.x
Subject(s) - tray , biology , plasmid , dna , binding site , dna binding protein , sequence (biology) , microbiology and biotechnology , genetics , transcription factor , gene , botany
Summary F plasmid traY protein binding to wild‐type or deleted regions containing the TraY‐binding site, sbyA , was studied in vitro. The principal DNA‐protein complex was formed with DNA segments including the sbyA site defined by footprinting and (with lesser affinity) with truncated segments that retained the leftward two‐thirds of sbyA. This located the major sequence determinants for TraY binding between bp 204 and 227 on the oriT map. For all sequences tested, bound TraY induced bending of approximateiy 50 to 55°, and centred between bp 214 and 221. Thermodynamic and mobility analyses indicated that two TraY protomers bind to sbyA. At higher TraY concentrations, additional TraY bound to the left of the sbyA in a region previously shown to bind IHF (site IHF A). TraY binding to this additional site ( sbyC ) was inhibited by IHF. Sequence similarities shared by sbyA, sbyB , and SbyC may include the critical base pairs for TraY binding.