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Nucleic Acids Research, 1977, Vol. 4, No. 8 2641-2654
© 1977

Articles

Action of pancreatic DNase: requirements for activation of DNA as a template-primer for DNA polymerase {propto}

E. Baril, J. Mitchener, L. Lee and B. Baril

Worcester Foundation for Experimental Biology, Inc. Shrewsbury, MA 01545, USA

Received April 18, 1977. Pancreatic DNase requires both Ca2+ and Mg2+ for its activity as measured by formation of an activated DNA template for in vitro DNA polymerase {alpha} assay and by the hyperchromic shift. Mn2+ can partially satisfy the Mg2+ requirement of the DNase for activation of DNA but the resulting template is only 50% as active in the DNA polymerase assay. When precautions are taken to avoid divalent ion contamination, pancreatic DNase is not active in the presence of Ca2+ or Mg2+ alone. Analysis of the DNA by sucrose gradient centrifugation shows that only in the presence of Ca2+ plus Mg2+ or Mn2+ does pancreatic DNase produce extensive strand breaks in the DNA. The activated DNA template that yields maximal DNA polymerase activity is low molecular weight material of 30,000 to 50,000 daltons.


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