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. 1974 May;71(5):1695–1700. doi: 10.1073/pnas.71.5.1695

Oncornavirus Expression in Human × Mouse Hybrid Cells Segregating Mouse Chromosomes

John D Minna *, Adi F Gazdar †, G Michael Iverson ‡, Thomas H Marshall *, Kurt Stromberg †, Samuel H Wilson †
PMCID: PMC388305  PMID: 4366059

Abstract

Human × mouse hybrid clones obtained by fusing transformed human (VA2) cells with embryonic mouse brain cells were tested for their ability to spontaneously express type C virus particles. It had been previously shown that these hybrid cells preferentially retained human chromosomes while mouse chromosomes were lost. The culture fluid from one cell line was found to contain type C particle markers in abundance, and typical budding C particles were observed in the cells by electron microscopy. In contrast, no particle markers were detected in the culture fluid from parental cells and several other hybrid cell lines. Subclones of the virus-positive cell line continued to lose mouse chromosomes and were found to vary more than 100-fold in their culture fluid DNA polymerase activity. The hybrid cell viruses, termed HMV1, banded in a sucrose gradient between 1.14 and 1.16 g/ml, possessed viral group-specific antigens, and exhibited B-tropic host range for replication in mouse embryo cells, but did not replicate in human cells when directly applied. The virus did not transform mouse cells but was able to rescue the defective murine sarcoma virus from sarcoma-positive, helper-virus-negative cells. Activity of the DNA polymerase associated with HMV1 was similar to the activity of Rauscher murine leukemia virus (MuLV) DNA polymerase in its preference for poly(rA) over poly(dA) as a template, use of endogenous template, detergent requirement, and inhibition by antiserum directed against MuLV.DNA polymerase. The results suggest that human × mouse hybrid cells segregating mouse chromosomes can spontaneously express endogenous type C viruses and that such hybrid cell lines may be used for the isolation of latent mammalian oncornaviruses and analysis of viral gene regulation.

Keywords: B-tropic virus, DNA polymerase, group-specific antigens, type C virus

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Selected References

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