A novel nuclear export activity in HIV-1 matrix protein required for viral replication

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Abstract

An important aspect of the pathophysiology of human immunodeficiency virus type-1 (HIV-1) infection is the ability of the virus to replicate in non-dividing cells1,2,3. HIV-1 matrix (MA), the amino-terminal domain of the Pr55 gag polyprotein (Pr55), bears a nuclear localization signal that promotes localization of the viral preintegration complex to the nucleus of non-dividing cells following virus entry3,4,5. However, late during infection, MA, as part of Pr55, directs unspliced viral RNA to the plasma membrane6, the site of virus assembly. How MA can mediate these two opposing targeting functions is not understood. Here we demonstrate that MA has a previously undescribed nuclear export activity. Although MA lacks the canonical leucine-rich nuclear export signal, nuclear export is mediated through the conserved Crmlp pathway and functions in both mammalian cells and yeast. A mutation that disrupts the MA nuclear export signal (MA-M4) mislocalizes Pr55 and genomic viral RNA to the nucleus, thereby severely impairing viral replication. Furthermore, we show that MA-M4 can act in a dominant-negative fashion to mislocalize genomic viral RNA even in the presence of wild-type MA. We conclude that the MA nuclear export signal is required to counteract the MA nuclear localization signal, thus ensuring the cytoplasmic availability of the components required for virion assembly.

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Figure 1: Characterization of a novel HIV-1 MA mutant.
Figure 2: Cellular distribution of viral and total poly(A)+ RNA by in situ hybridization.
Figure 3: Cellular distribution of Pr55 and MA variants.
Figure 4: Nuclear export activity of HIV-1 MA.
Figure 5: Analysis of a MA NES-NLS double mutant.
Figure 6: Mislocalization of RNA by Pr55-M4 requires the nucleocapsid-Psi site interaction.

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Acknowledgements

We thank J. Mondor, N. Bakker and L. Ross for manuscript preparation. We also thank M. Yoshida for generous donation of Leptomycin B, and M. Rosbash for yeast strains. We thank J. Kan, J. Teodoro, M. Sharkey, J.-M. Jacque, B. Brichacek, S. Swingler and M. Zapp for scientific discussions. This work was supported in part by grants from the NIH to M.R.G. and M.S., and A.B. is supported in part by a Fogarty International Research Collaboration Award. M.R.G. is an investigator of the Howard Hughes Medical Institute.

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Correspondence to Michael R. Green.

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Dupont, S., Sharova, N., DéHoratius, C. et al. A novel nuclear export activity in HIV-1 matrix protein required for viral replication. Nature 402, 681–685 (1999) doi:10.1038/45272

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