Albany 2015:Book of Abstracts
June 9-13 2015
©Adenine Press (2012)
Influence of 1,3-diazaadamantane Derivative on the Strand Exchange between Short Oligonucleotides
Adamantane derivatives are known for broad spectrum of applications in medicine and bioscience due to the large size and conformational rigidity of the adamantane backbone. Owing to the presence of two ternary nitrogen atoms in the backbone of the 1,3-diazaadamantanes, these conjugates of inert adamantine acquire a chemical activity and an ability to be isolated in a water-soluble hydrochloride form. As we reported earlier, among a number of characterized 1,3-diazaadamantane derivatives that stimulate DNA Strand Exchange Reaction (SER), the compound HG122 appeared to be most promising for further study ( Gabrielian et al.,2013., Mamaeva et.al.,2014).Here we present the results of the SER titration via an excess of competing oligonucleotide, according to ( Reynaldo et.al., 2000). The sensitivity of the HG122 promoted SER to limited heterology between the reaction substrates was also estimated. As seen from the Figure, both HG122 and Rec A protein promote SER. In both cases, SER exhibits decreased sensibility to homology violation unlike SER promoted by other agents, such as linker histone H1, or mammalian meiotic system protein Hop2. The latter exhibit the same sensitivity to heterology as that of spontaneous SER at increased temperature (Bocharova et.al..2012, Pezza et.al., 2014).Thus, 1,3-diazaadamantane derivative HG122 acts similar to recombinase Rec A family. This ligand facilitates SER, destabilizes the DNA duplex and appreciably reduces SER sensitivity to heterology between SER substrates. Therefore, compounds of this class are worthy models for further study of different aspects of SER mechanisms.
Figure: Kinetic curves of SER between completely homological substrates (perf) and substrates, containing singular interruption (mm), introduced by one base replacement in the central part of single-stranded oligonucleotide, (A) - in presence of HG122 and spontaneous SER in the absence of HG122 at increased temperature (Spont), (B)- in presence of Rec A protein. The reaction mixture contained equimolar amounts of double-and fluorescent single-stranded substrate ( Bocharova et.al.,2012).
Mamaeva O.K., Gabrielian A., Harutjunyan G., Bocharova T.N., Smirnova E.A., Volodin A.A., Shchjolkina A.K., Kaljuzhnij D.N. (2014). Molecular Biology, 48, 5, 741-748.
Reynaldo L.P., Vologodskij A.V., Neri B.P., Lyamichev V.I. (2000) J.Mol.Biol, 297, 511-520.
Bocharova T.N., Smirnova E.A., Volodin A.A. (2012). Biopolymers, 97, 229-239.
Pezza R.J., Voloshin O.N., Volodin A.A., Boateng K.A., Bellani M.A., Mazin A.V., Camerini-Otero R.D.(20014). Nucleic Acids Research, 42, 4, 2346- 2357.
1Research-technological Center of Organic & Pharmacological Chemistry (Armenian National Academy of Sciences)