Acquisition of new proviral copies in avian lymphoid cells transformed by reticuloendotheliosis virus.

AUTOR(ES)

Zhang, J Y

RESUMO

The expression of the v-rel oncogene of avian reticuloendotheliosis virus (REV-T) transforms and immortalizes very immature avian lymphoid cells. In REV-T-transformed lymphoid cells which were persistently infected with reticuloendotheliosis-associated virus (REV-A), the REV-T proviral copy number increases after the initial integration event. In 23 independently derived REV-T-transformed cell lines, 15 of the 18 virus-producing cell lines had acquired additional proviruses. The rate at which the newly acquired proviral sequences accumulated differed for various cell lines. In some cell lines, additional REV-T proviral copies could be detected as early as 8 months after the initial integration event. A correlation exists between the number of REV-T proviral sequences and the length of time which a given cell line had been propagated in culture. The integration sites occupied by the newly acquired REV-T proviruses were distinct. In contrast, reticuloendotheliosis-associated virus proviral sequences in these REV-T-transformed virus-producing lymphoid cells did not increase during in vitro culture. Furthermore, the acquisition of additional REV-T proviral sequences did not occur in non-virus-producing cell lines. Two of the newly acquired proviral sequences were molecularly cloned and analyzed by restriction endonuclease mapping. Although the newly acquired REV-T proviruses have not sustained major deletions, the viral sequences and the v-rel oncogene display numerous restriction enzyme polymorphisms. The cellular flanking sequences of two newly acquired REV-T proviruses analyzed were unique and shared no homology with flanking sequences of the other REV-T proviruses in these transformed cells. The nucleotide sequence of the virus-cellular DNA junctions of one newly acquired provirus and its cellular sequence prior to proviral integration were defined. A 5-base-pair direct repeat of cellular origin was present on each side of the long terminal repeat, indicating that the mechanism of acquisition of additional REV-T proviral sequences used reverse transcription and integration of new REV-T proviral copies.

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