Isolation and characterization of polyoma uncoating intermediates from the nuclei of infected mouse cells

Abstract

A method was developed which enabled the efficient recovery of polyoma uncoating intermediates from the nuclei of infected cells at early times after infection (15 min to 12 h). Cells were infected with radiolabeled virus and lysed with the detergent Nonidet P-40. The nuclei were then collected and sonicated, and the products were analyzed on sucrose gradients. The uncoating intermediate sedimented at 190S and was a viral DNA-protein complex closely associated with a structure of host origin. The host material associated with the 190S uncoating intermediate was determined by polyacrylamide gel electrophoresis and visualized by electron microscopy. The amount of 190S uncoating intermediate found in the nucleus increased with time after infection. The viral DNA was predominantly for I. All of the viral proteins were present in the 190S uncoating intermediate in amounts similar to those found in viral DNA-protein complex cores.