title: Targeting Holliday junctions by origin DNA-binding protein of herpes simplex virus type 1.
abstract: In the present paper, the interactions of the origin binding protein (OBP) of herpes simplex virus type 1 (HSV1) with synthetic four-way Holliday junctions (HJs) were studied using electrophoresis mobility shift assay and the FRET method and compared with the interactions of the protein with duplex and single-stranded DNAs. It has been found that OBP exhibits a strong preference for binding to four-way and three-way DNA junctions and possesses much lower affinities to duplex and single-stranded DNAs. The protein forms three types of complexes with HJs. It forms complexes I and II which are reminiscent of the tetramer and octamer complexes with four-way junction of HJ-specific protein RuvA of Escherichia coli. The binding approaches saturation level when two OBP dimers are bound per junction. In the presence of Mg2+ions (≥2 mM) OBP also interacts with HJ in the stacked arm form (complex III). In the presence of 5 mM ATP and 10 mM Mg2+ions OBP catalyzes processing of the HJ in which one of the annealed oligonucleotides has a 3'-terminal tail containing 20 unpaired thymine residues. The observed preference of OBP for binding to the four-way DNA junctions provides a basis for suggestion that OBP induces large DNA structural changes upon binding to Box I and Box II sites in OriS. These changes involve the bending and partial melting of the DNA at A+T-rich spacer and also include the formation of HJ containing Box I and Box II inverted repeats and flanking DNA sequences.
authors.affiliation: a Engelhardt Institute of Molecular Biology , Russian Academy of Sciences , ul. Vavilova 32, 119991 Moscow , Russia.
authors.forename: E D
authors.initials: ED
authors.lastname: Moiseeva
authors.affiliation: a Engelhardt Institute of Molecular Biology , Russian Academy of Sciences , ul. Vavilova 32, 119991 Moscow , Russia.
authors.forename: N P
authors.initials: NP
authors.lastname: Bazhulina
authors.affiliation: b Russian Cardiology Research-and-Production Complex , 3ya Cherepkovskaya ul. 15a, 121552 Moscow , Russia.
authors.forename: Y G
authors.initials: YG
authors.lastname: Gursky
authors.affiliation: a Engelhardt Institute of Molecular Biology , Russian Academy of Sciences , ul. Vavilova 32, 119991 Moscow , Russia.
authors.forename: S L
authors.initials: SL
authors.lastname: Grokhovsky
authors.affiliation: a Engelhardt Institute of Molecular Biology , Russian Academy of Sciences , ul. Vavilova 32, 119991 Moscow , Russia.
authors.forename: A N
authors.initials: AN
authors.lastname: Surovaya
authors.affiliation: a Engelhardt Institute of Molecular Biology , Russian Academy of Sciences , ul. Vavilova 32, 119991 Moscow , Russia.
authors.forename: G V
authors.initials: GV
authors.lastname: Gursky
mesh_terms: D001483:Base Sequence; D004261:DNA Replication; D045566:DNA, Cruciform / Q000037:antagonists & inhibitors*; D004279:DNA, Viral / Q000235:genetics* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000494:pharmacology*; D024202:Electrophoretic Mobility Shift Assay; D018259:Herpesvirus 1, Human; D006801:Humans; D009690:Nucleic Acid Conformation
chemical_list: D045566:DNA, Cruciform; D004279:DNA, Viral; D004268:DNA-Binding Proteins
keywords: ATP; E. coli RuvA protein; four-way DNA Holliday junction; herpes simplex virus type 1; viral origin DNA-binding protein