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Journal of Virology, February 2008, p. 1600-1604, Vol. 82, No. 3
0022-538X/08/$08.00+0     doi:10.1128/JVI.02295-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Gag-Processing Defect of Human Immunodeficiency Virus Type 1 Integrase E246 and G247 Mutants Is Caused by Activation of an Overlapping 5' Splice Site{triangledown}

Dibyakanti Mandal, Zehua Feng, and C. Martin Stoltzfus*

Department of Microbiology, University of Iowa, Iowa City, Iowa 52242

Received 22 October 2007/ Accepted 5 November 2007

We have previously described several human immunodeficiency virus type 1 (HIV-1) mutants that are characterized by an excessive-RNA-splicing phenotype and reduced virus particle production. In one of these mutants (NLD2up), the sequence of 5' splice site D2 was changed to a consensus splice donor site. This splice site overlaps the HIV-1 integrase reading frame, and thus, the NLD2up mutant also bears a G-to-W change at amino acid 247 of the integrase. A previously described E-to-K mutant at position 246 of the C-terminal domain of the integrase, which resulted in a G-to-A mutation at the +3 position of overlapping splice donor D2 (NLD2A3), was also shown to affect virus particle production and Gag protein processing. By using second-site mutations to revert the excessive-splicing phenotype, we show that the effects on Gag protein processing and virus particle production of both the NLD2up and NLD2A3 mutants are caused by excessive viral RNA splicing due to the activation of the overlapping 5' splice site and not to the changes in the integrase protein. Both integrase protein mutations, however, are lethal for virus infectivity. These studies suggest that changes in the usage of overlapping splice sites may be a possible alternative explanation for a defective virus phenotype resulting from changes in protein-coding sequences or in the nucleotide sequence during codon optimization.

* Corresponding author. Mailing address: Department of Microbiology, University of Iowa, Iowa City, IA 52242. Phone: (319) 335-7793. Fax: (319) 335-9006. E-mail: marty-stoltzfus{at}uiowa.edu

{triangledown} Published ahead of print on 21 November 2007.

Journal of Virology, February 2008, p. 1600-1604, Vol. 82, No. 3
0022-538X/08/$08.00+0     doi:10.1128/JVI.02295-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Mandal, D., Exline, C. M., Feng, Z., Stoltzfus, C. M. (2009). Regulation of vif mRNA Splicing by Human Immunodeficiency Virus Type 1 Requires 5' Splice Site D2 and an Exonic Splicing Enhancer To Counteract Cellular Restriction Factor APOBEC3G. J. Virol. 83: 6067-6078 [Abstract] [Full Text]  
  • Exline, C. M., Feng, Z., Stoltzfus, C. M. (2008). Negative and Positive mRNA Splicing Elements Act Competitively To Regulate Human Immunodeficiency Virus Type 1 Vif Gene Expression. J. Virol. 82: 3921-3931 [Abstract] [Full Text]  


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