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Nucleic Acids Research, 1978, Vol. 5, No. 5 1715-1728
© 1978

Articles

DNA polymerase {alpha} is associated with replicating SV4O nucleoprotein complexes*

Bernd Otto and Ellen Fanning

Fachbereich Biologie, University of Konstanz D-7750 Konstanz, GFR

Received March 23, 1978.

Simian virus 40 (SV40) nucleoprotein complexes were extracted from nuclei of infected monkey cells and fractionated on neutral sucrose density gradients. Complexes which contai replicating SV40 DNA (95S) separated well from those containing closed circular supercoiled viral DNA (75S). DNA polymerase activity was associated with the replicating nucleoprotein complexes but not with the slower sedimenting complexes. This DNA polymerase activity coprecipitated with the nucleoprotein complexes in the presence of MgCl2 and remained associated with the 95S complexes. This DNA polymerase activity has been identified as primarily DNA polymerase {alpha} on the basis of its sedimentation behavior, optimum salt concentration, and sensitivity to N-ethylmaleimide. DNA polymerase {gamma} activity was also detected in the complexes, but DNA polymerase ß was not associated with the complexes.

*Part of this work was presented at the NATO Advanced Study Institute on "DNA Synthesis, Present and Future", Palermo, Italy, June 20–29, 1977.


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