Analysis of HIV-2 Vpx by modeling and insertional mutagenesis

Lisa A. Mahnke, Michael A. Belshan, Lee Ratner

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Vpx facilitates HIV-2 nuclear localization by a poorly understood mechanism. We have compared Vpx to an NMR structure HIV-1 Vpr in a central helical domain and probed regions of Vpx by insertional mutagenesis. A predicted loop between helices two and three appears to be unique, overlapping with a known novel nuclear localization signal. Overall, Vpx was found to be surprisingly flexible, tolerating a series of large insertions. We found that insertion within the polyproline-containing C-terminus destabilizes nuclear localization, whereas mutating a second helix in the central domain disrupts viral packaging. Other insertional mutants in the predicted loop and in a linker region between the central domain and the C-terminus may be useful as sites of intramolecular tags as they could be packaged adequately and retained preintegration complex associated integration activity in a serum starvation assay. An unexpected result was found within a previously defined nuclear localization motif near aa 71. This mutant retained robust nuclear localization in a GFP fusion assay and was competent for preintegration complex associated nuclear import. In summary, we have modeled helical content in Vpx and assessed potential sites of intramolecular tags which may prove useful for protein-protein interactions studies.

Original languageEnglish
Pages (from-to)165-174
Number of pages10
JournalVirology
Volume348
Issue number1
DOIs
StatePublished - Apr 25 2006
Externally publishedYes

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HIV-2
Insertional Mutagenesis
Virus Assembly
Nuclear Localization Signals
Cell Nucleus Active Transport
Starvation
HIV-1
Proteins
Serum
polyproline

All Science Journal Classification (ASJC) codes

  • Virology
  • Infectious Diseases

Cite this

Analysis of HIV-2 Vpx by modeling and insertional mutagenesis. / Mahnke, Lisa A.; Belshan, Michael A.; Ratner, Lee.

In: Virology, Vol. 348, No. 1, 25.04.2006, p. 165-174.

Research output: Contribution to journalArticle

Mahnke, Lisa A. ; Belshan, Michael A. ; Ratner, Lee. / Analysis of HIV-2 Vpx by modeling and insertional mutagenesis. In: Virology. 2006 ; Vol. 348, No. 1. pp. 165-174.
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