Properties of Intracytoplasmic A Particles Isolated from Oncornavirus-Producing Human Cells

Author:

Bukrinskaya A. G.1,Miller G. G.1,Lebedeva E. N.1,Agaphonova L. V.1,Masurenko N. N.1,Ilyin K. V.1

Affiliation:

1. The D. I. Ivanovsky Institute of Virology and the N. F. Gamaleya Institute of Microbiology and Epidemiology, Academy of Medical Sciences, Moscow, U. S. S. R.

Abstract

A particles with the diameter of 70 to 80 nm were isolated from the cytoplasm of HEp-2, HeLa, and AO cells producing oncornavirus of Mason-Pfizer-like type. Most of the A particles banded at 1.23 to 1.24 g/ml, whereas 3 to 10% banded at 1.29 g/ml in equilibrium sucrose gradients. They banded at 1.30 g/ml in CsCl gradients suggesting that they contained 8% RNA. Individual A particles sedimented at 200 to 250 S in velocity sucrose gradients, but their significant part was found aggregated and sedimented at more than 300 S . They were resistant to RNase digestion. A particles possessed polymerase activity which was preferentially activated by Mn 2+ rather than by Mg 2+ , the RNA template being 60 S RNA. Cross-hybridization with two DNA products and immunoassay showed that A particles and Mason-Pfizer-like oncornavirus produced by the same cells contained neither homological RNA sequences nor common antigens, suggesting that A particles are not intracellular precursors of Mason-Pfizer-like oncornavirus but represent an independent oncornavirus. Hybridization of A particle RNA with excess of cellular DNA revealed about 20 proviral copies per HEp-2 cell genome and no proviral copies in human embryo and placenta cell genomes.

Publisher

American Society for Microbiology

Subject

Virology,Insect Science,Immunology,Microbiology

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