Differences between the endogenous and exogenous dna sequences of rous-associated virus-O

Abstract

DNA sequences related to the endogenous retrovirus of chickens, Rous-associated virus-0 (RAV-0), have been examined using site-specific DNA endonuclease analysis of cellular DNA derived from line 15 and line 100 chickens. Individual embryos from both inbred lines were used as a source of embryonic fibroblasts from which cellular DNA was isolated. Analysis of DNA containing either endogenous RAV-0 sequences alone or both endogenous and exogenous RAV-0 sequences produced identical patterns of RAV-0-specific DNA fragments after digestion with the endonucleases Eco RI, Hind III, Bgl II, Bam HI or Xho I. Similar analysis with endonucleases Hinc II or Hha I, however, produced several RAV-0-specific DNA fragments which were derived from cellular DNA containing both endogenous and exogenous RAV-0 sequences but not from cellular DNA containing only endogenous sequences. Although some differences exist between the DNA fragments specific for the endogenous viral sequences of line 15 and line 100 cellular DNA, the DNA fragments specific for the exogenous viral sequences were identical between the two inbred lines. Cleavage of an unintegrated linear RAV-0 DNA molecule with Hinc II or Hha I produced DNA fragments identical to those specific for the exogenously acquired RAV-0 provirus. This suggests that these characteristic fragments contain no cellular DNA. The potential DNA junction fragments containing both viral and cellular DNA, identified after analysis of DNA that contains both endogenous and exogenous viral sequences, were identical to those observed after analysis of DNA containing only endogenous viral sequences. These results support the following conclusions. First, exogenous proviral sequences are integrated into chicken cell DNA following an interaction between viral and cellular DNA that is specific with respect to the virus and nonspecific with respect to the cell. Second, both the free linear RAV-0 DNA intermediate and the newly integrated exogenous provirus contain specific endonuclease sites that are not found in endogenous RAV-0 DNA sequences. These results suggest that the formation of the exogenous DNA provirus involves specific alteration of the endogenous viral DNA sequences before reinsertion of the sequences as the exogenous RAV-0 DNA provirus. It is possible that newly integrated exogenous RAV-0 sequences are characterized by specific differences in the pattern of base methylation and a limited sequence arrangement.