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Journal of Virology, January 2008, p. 503-512, Vol. 82, No. 1
0022-538X/08/$08.00+0     doi:10.1128/JVI.01441-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Rous Sarcoma Virus (RSV) Integration In Vivo: a CA Dinucleotide Is Not Required in U3, and RSV Linear DNA Does Not Autointegrate{triangledown}

Jangsuk Oh,1 Kevin W. Chang,1 Rafal Wierzchoslawski,1 W. Gregory Alvord,2 and Stephen H. Hughes1*

HIV Drug Resistance Program,1 Data Management Services, National Cancer Institute at Frederick, Frederick, Maryland 21702-12012

Received 2 July 2007/ Accepted 15 October 2007

The sequences required for integration of retroviral DNA have been analyzed in vitro. However, the in vitro experiments do not agree on which sequences are required for integration: for example, whether or not the conserved CA dinucleotide in the 3' end of the viral DNA is required for normal integration. At least a portion of the problem is due to differences in the experimental conditions used in the in vitro assays. To avoid the issue of what experimental conditions to use, we took an in vivo approach. We made mutations in the 5' end of the U3 sequence of the Rous sarcoma virus (RSV)-derived vector RSVP(A)Z. We present evidence that, in RSV, the CA dinucleotide in the 5' end of U3 is not essential for appropriate integration. This result differs from the results seen with mutations in the U5 end, where the CA appears to be essential for proper integration in vivo. In addition, based on the structure of circular viral DNAs smaller than the full-length viral genome, our results suggest that there is little, if any, integrase-mediated autointegration of RSV linear DNA in vivo.

* Corresponding author. Mailing address: HIV Drug Resistance Program, NCI at Frederick, P.O. Box B, Bldg. 539, Rm. 130A, Frederick, MD 21702-1201. Phone: (301) 846-1619. Fax: (301) 846-6966. E-mail: Hughes{at}ncifcrf.gov

{triangledown} Published ahead of print on 24 October 2007.

Journal of Virology, January 2008, p. 503-512, Vol. 82, No. 1
0022-538X/08/$08.00+0     doi:10.1128/JVI.01441-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Oh, J., Chang, K. W., Hughes, S. H. (2008). Integration of Rous Sarcoma Virus DNA: a CA Dinucleotide Is Not Required for Integration of the U3 End of Viral DNA. J. Virol. 82: 11480-11483 [Abstract] [Full Text]  
  • Chang, K. W., Oh, J., Alvord, W. G., Hughes, S. H. (2008). The Effects of Alternate Polypurine Tracts (PPTs) and Mutations of Sequences Adjacent to the PPT on Viral Replication and Cleavage Specificity of the Rous Sarcoma Virus Reverse Transcriptase. J. Virol. 82: 8592-8604 [Abstract] [Full Text]  


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