Effects of intercalators on complexation of RecA with duplex DNA
Artikel i vetenskaplig tidskrift, 1995

To elucidate the binding mode of recombination protein A (RecA) to double-stranded (ds) DNA, the effects on the RecA-DNA interaction of several mono- and bisintercalators of the acridine, phenanthridine, and cyanine classes have been investigated by linear dichroism spectroscopy. Simple monointercalators lacking side chains efficiently promoted the binding of RecA to dsDNA in the absence of nucleotide cofactor, which is otherwise required. Bisintercalators varied in their ability to induce RecA binding, while monointercalators with aminoalkyl side chains proved inefficient. Modification of DNA structure by the intercalator appears to be necessary for induction of RecA binding, but if the intercalator has a bulky minor-groove-binding side chain, it does not induce RecA binding. In detailed studies with acridines, neither the binding geometry of intercalators nor the structure of DNA was significantly modified upon binding of RecA without cofactor. Judged by circular dichroism, similar ReA conformational changes accompanied bis-9-aminoacridine- and ATPyS-induced RecA association with DNA. In the presence of ATPyS, the intercalators inhibited the rate of RecA binding to dsDNA and were extruded from DNA upon binding of RecA. This competitive aspect may suggest that intercalation of some amino acid residue(s) plays a role in nucleotide-induced RecA binding. The stoichiometry of the RecA-DNA-intercalator filament was determined; in the fully formed filament the base pair:intercalator ratio is 2, and the base pair:RecA ratio also 2. This contrasts with a base pair:RecA ratio of 3 in the ATPyS-induced filament, although in both cases the DNA experiences 50% extension.

angle

protein

binding

linear dichroism spectroscopy

neutron-scattering

cofactor

electron-microscopy

double-stranded dna

spectroscopy

fluorescence

nucleic-acids

escherichia-coli

Författare

Eimer Tuite

Institutionen för fysikalisk kemi

S. K. Kim

Bengt Nordén

Institutionen för fysikalisk kemi

M. Takahashi

Biochemistry

0006-2960 (ISSN) 1520-4995 (eISSN)

Vol. 34 16365-16374

Ämneskategorier

Biokemi och molekylärbiologi

DOI

10.1021/bi00050a017