A research collaboration is proposed between the multidisciplinary genosensor technology development team at the Houston Advanced Research Center (HARC) and the Laboratory of Nucleoproteins and Proteins at the Zelinsky Institute of Organic Chemistry in Moscow, to apply DNA chip technology in the analysis of interactions between nucleic acids and polyamines and proteins. An array of oligonucleotides, attached to glass or gold surfaces at one end (DNA chip, or genosensor), will be fabricated (at HARC, under the parent grant) and used to simultaneously analyze the binding of biological polyamines and ribosomal proteins to numerous oligonucleotide sequences. Detection of binding will be performed by fluorography (or autoradiography) and by use of microfabricated electronic (permittivity) and charge coupled device (CCD) chips, supplied by the HARC team. The experiments will investigate the influence of nucleotide sequence the secondary structure on the kinetic and equilibrium constants for binding of several biological polyamines to nucleic acids. The results of the polyamine studies may reveal information about the in vivo function of natural polyamines, such as their role in the regulation of cellular proliferation. Experiments with ribosomal proteins and oligonucleotide sequences found in ribosomal RNA will characterize the complex interactions within the ribosome. The latter work is expected to produce valuable experience to both the Zelinsky and HARC research teams concerning the use of DNA chips for studies of protein-nucleic acid interactions. The proposed FIRCA project will help the Budowsky group maintain a world leadership position in the field of nucleic acid-protein interactions, and should produce results which will enable acquisition of further NIH funding for expansion of the collaborative program.
