Alteration of location of dimer linkage sequence in retroviral RNA: Little effect on replication or homologous recombination

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Output type: Journal article

Author list: Jones J., Allan R., Temin H.

Publisher: American Society for Microbiology

Publication year: 1993

Journal: Journal of Virology (0022-538X)

Volume number: 67

Issue number: 6

Start page: 3151

End page: 3158

Number of pages: 8

ISSN: 0022-538X

eISSN: 1098-5514

URL: http://api.elsevier.com/content/abstract/scopus_id:0027191916

Abstract

Retrovirus particles contain a dimer of retroviral genomic RNA. A defined region of the retrovirus genome has previously been shown to be important for both dimerization and encapsidation. To study the importance of the position of this encapsidation and dimerization signal for retroviral replication and homologous recombination, we used a previously described spleen necrosis virus-based helper cell system. This system allows retroviral vectors with multiple genetic markers to be studied after a single cycle of retroviral replication. The sequence responsible for dimerization, the encapsidation/dimer linkage sequence (E/DLS), was moved from its normal location near the 5′ end of the retroviral genome to a location near the 3′ end of the genome. We characterized four pairs of retroviral vectors: (i) with both E/DLSs at the 5′ ends of the genomes, (ii) with both E/DLSs at the 3′ ends of the genomes, and (iii) two with one E/DLS at the 5′ end of the genome and one at the 3′ end of the genome. We found that moving the E/DLS to the 3′ end of the genome resulted in at most an approximately factor of 5 reduction in virus titer in a single cycle of retroviral replication. Furthermore, we found no changes that were attributable to the alteration of the position of the E/DLS in the minus-strand DNA primer transfers or the plus-strand DNA primer transfers, the rate of homologous recombination, or the number of internal template switches in recombinant proviruses. These results indicate that any alignment or conformation necessary for retroviral replication or recombination is not the result of the position of the E/DLS.

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