Analysis of retroviral proteins by two-step DNA, zinc binding, and modified Southern-western blotting

Using Southern-Western and zinc-blotting techniques, the interactions of single-stranded DNA and zinc with structural proteins of type B, C, and D retroviruses were examined. Besides nucleic capsid proteins of retroviruses known to form complexes with nucleic acids, some other core proteins of type C retroviruses were shown to bind nucleic acids. All nucleic capsid proteins of the examined retroviruses appeared to be also zinc binding. In the present report, we propose two protein-blotting techniques that could be performed on a single nitrocellulose membrane. A first technique allows to detect zinc- and DNA-binding proteins immobilized on the membrane; a second (a modification of Southern-Western blotting) makes it possible to detect DNA-binding proteins followed by immunological reprobing.