Monoclonal antibody that blocks phosphoinositide-dependent activation of mouse tumor DNA polymerase alpha |
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| Authors: | V L Sylvia J O Norman G M Curtin D L Busbee |
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| Institution: | 1. Department of Anatomy , College of Veterinary Medicine, Texas A&M University, College Station, TX 77843, USA;2. Department of Physiology and Pharmacology, College of Veterinary Medicine, Texas A&M University, College Station, TX 77843, USA;3. Veterinary Toxicology and Entomology Research Laboratory USDA, ARS, College Station, TX 77841, USA;1. Third Faculty of Medicine, Charles University, Prague, Czech Republic;2. Department of Ophthalmology, University Hospital Královské Vinohrady, Prague, Czech Republic;3. Department of Ophthalmology, University Hospital Hradec Králové, Hradec Králové, Czech Republic;4. Department of Ophthalmology, Faculty of Medicine, Masaryk University, Brno, Czech Republic;5. Department of Ophthalmology, University Hospital Brno, Brno, Czech Republic;6. Department of Ophthalmology, University Hospital Ostrava, Ostrava, Czech Republic;7. Department of Internal Medicine, University Hospital Královské Vinohrady, Prague, Czech Republic |
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| Abstract: | A monoclonal antibody (MaB) against mouse sarcoma DNA polymerase alpha was isolated from the culture medium of an IgG-secreting hybridoma. The MaB demonstrated reactivity against two murine DNA polymerase alpha preparations and a calf thymus DNA polymerase alpha. Murine sarcoma polymerase was activated in vitro by phosphatidylinositol-4-monophosphate (PIP) showing increased deoxynucleotidyltransferase activity and enhanced binding affinity to activated DNA template. The MaB did not neutralize polymerase activity, but blocked further activation of the enzyme by PIP. Treatment of polymerase with MaB prior to treatment with PIP inhibited both increased enzyme activation and increased binding of the enzyme to DNA template. Treatment of polymerase with MaB subsequent to treatment with PIP did not block enzyme activation or increased DNA template binding. The data suggest that this anti-DNA polymerase alpha IgG is directed against a regulatory subunit of the polymerase rather than the catalytic subunit. The antibody may serve to distinguish between DNA polymerase alpha preparations with distinctly different regulatory subunits. |
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