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Inhibition of HIV replication by dominant negative mutants of Sam68, a functional homolog of HIV-1 Rev

An Erratum to this article was published on 01 July 1999

Abstract

The HIV-1 Rev protein facilitates the nuclear export of mRNA containing the Rev response element (RRE) through binding to the export receptor CRM-1. Here we show that a cellular nuclear protein, Sam68 (Src-associated protein in mitosis), specifically interacts with RRE and can partially substitute for as well as synergize with Rev in RRE-mediated gene expression and virus replication. Differential sensitivity to leptomycin B, an inhibitor of CRM-1, indicates that the export pathways mediated by Rev and Sam68 are distinct. C-terminally deleted mutants of Sam68 inhibited the transactivation of RRE-mediated expression by both wild-type Sam68 and Rev. They were retained in the cytoplasm and impeded the nuclear localization of Rev in co-expressed cells. These mutants also inhibited wild-type HIV-1 replication to the same extent as the RevM10 mutant, and may be useful as anti-viral agents in the treatment of AIDS.

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Figure 1: Effects of wild-type and mutant Sam68 on RRE-mediated CAT expression.
Figure 2: Microinjection of Sam68Δ96 expression plasmid activates RRE-mediated reporter gene expression in human cells.
Figure 3: Interaction of Sam68 with RRE in vitro and in vivo.
Figure 4: Sam68 synergizes with Rev in RRE directed reporter gene expression and HIV-1 expression.
Figure 5: C-terminally deleted mutants of Sam68 inhibit Rev and Sam68 function.
Figure 6: In vitro binding of transdominant mutant Sam68C'Δ330-443 to RRE and Rev.
Figure 7: Co-localization of Rev and Sam68 with transdominant mutant C'Δ330-443.

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Acknowledgements

We also thank D. Looney, C. Barroga and K. Kuhen for critical review of the manuscript, and C. Westberg for editorial assistance. Recombinant Rev was obtained through the AIDS Research and Reference Reagents Program, Division of AIDS, National Institute of Allergy and Infectious Diseases (D. Rekosh, M.-L. Hammerskjold and M. Orsini). This work was supported in part by NIH grant GM056089 to F.W.-S. We also acknowledge the support of the Center for AIDS Research (NIH Grant P30AI 36214-06) and National Center for Microscopy and Imaging Research at UCSD (NIH grant # RR04050; PI, M. H. Ellisman).

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Reddy, T., Xu, W., Mau, J. et al. Inhibition of HIV replication by dominant negative mutants of Sam68, a functional homolog of HIV-1 Rev. Nat Med 5, 635–642 (1999). https://doi.org/10.1038/9479

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