Rose Bengal mediated inhibition of DNA polymerases. 2. Mechanism of inhibition of avian myeloblastosis virus reverse transcriptase under photooxidative conditions

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Abstract
The primary effect of photooxidation on the enzyme-Rose Bengal complex was the loss of template-primer binding ability within 5 min of exposure to light. The presence of template-primer, but not the substrate deoxynucleotides, consistently provided partial protection against Rose Bengal mediated photooxidative inactivation. Preformed enzyme-template-primer complexes were not dissociated by Rose Bengal under these conditions. Prolonged exposure (25-30 min) of this complex to light in the presence of Rose Bengal led to complete inactivation of catalytic activity without affecting the ability of enzyme to bind to template-primer. Thus, oxidative inactivation of avian myeloblastosis virus reverse transcriptase in the presence of Rose Bengal involves a domain within the enzyme that contains the template-primer binding site and an additional site which is required for the expression of the polymerization and nuclease activities of the reverse transcriptase.

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