Aggregation of RecA-derived peptides on single-stranded oligonucleotides triggered by schiff base-mediated crosslinking.
We here show that single-stranded oligonucleotides containing 5-formyl-2'-deoxyuridine (fdU) can crosslink the peptides derived from the DNA binding site of RecA protein through a Schiff base formation. The ability of crosslinking of fdU-containing oligonucleotides was investigated using a series of peptides whose amino acid residues spanning the center of the RecA-derived peptide were sequentially replaced with lysine. Circular dichroism (CD) spectroscopy, gel mobility shift assay and sedimentation experiment demonstrated that crosslinking reaction proceeded efficiently only when the peptides bound to the oligonucleotides.
Sugiyama
Toru
T
Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Japan. csugi@mail.ecc.u-tokyo.ac.jp
Kittaka
Atsushi
A
Takayama
Hiroaki
H
Tomioka
Mitsugu
M
Ida
Yoshiteru
Y
Kuroda
Reiko
R
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D002498:Centrifugation / Q000379:methods; D002942:Circular Dichroism; D003432:Cross-Linking Reagents / Q000737:chemistry*; D004277:DNA, Single-Stranded / Q000737:chemistry*; D003857:Deoxyuridine / Q000031:analogs & derivatives* / Q000737:chemistry; D024202:Electrophoretic Mobility Shift Assay; D004926:Escherichia coli / Q000201:enzymology; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry*; D011938:Rec A Recombinases / Q000737:chemistry*; D012545:Schiff Bases / Q000737:chemistry*
D003432:Cross-Linking Reagents; D004277:DNA, Single-Stranded; D009841:Oligonucleotides; D010446:Peptide Fragments; D012545:Schiff Bases; C061519:5-formyl-2'-deoxyuridine; D011938:Rec A Recombinases; D003857:Deoxyuridine