Reticuloendotheliosis virus-transformed cells contain infectious and noninfectious proviral sequences in different chromosomes

Abstract

The integration site of reticuloendotheliosis virus proviral DNA in the DNA from a cloned reticuloendotheliosis virus-transformed bone marrow cell line was studied. These reticuloendotheliosis virus-transformed bone marrow cells produce a replication-defective transforming virus (REV-T) and a nontransforming helper virus, designated reticuloendotheliosis-associated virus (REV-At). The DNA from this REV-T transformed bone marrow cell line was hybridized with 3H-labeled in vitro synthesized DNA complementary to the reticuloendotheliosis virus (RE) genome to show that these cells contain approximately five genome equivalents per haploid genome. These hybridization experiments did not distinguish between the transforming and nontransforming virus sequences in these cells. Metaphase chromosomes have been isolated from colcemid-treated bone marrow cells and separated into four different size classes on zonal gradients. Hybridization experiment with RE-specific cDNA indicated that all four size classes of chromosomes contain RE-specific sequences. To locate the infectious provirus DNA of the helper virus REV-At, transfection experiments were performed with DNA extracted from the chromosomes of each size class. Infectious REV-At provirus DNA was found principally in one size class. This size class contained the intermediate size macrochromosomes 5 through 11. These results suggest that REV-At proviral DNA must be integrated into a specific chromosome to produce infectious virus.