The nucleocapsid domain of gag is dispensable for actin incorporation into HIV-1 and for association of viral budding sites with cortical F-actin

Sarah Stauffer, Sheikh Abdul Rahman, Alex de Marco, Lars Anders Carlson, Bärbel Glass, Heike Oberwinkler, Nikolas Herold, John A.G. Briggs, Barbara Müller, Kay Grünewald, Hans Georg Kräusslich

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Abstract

Actin and actin-binding proteins are incorporated into HIV-1 particles, and F-actin has been suggested to bind the NC domain in HIV-1 Gag. Furthermore, F-actin has been frequently observed in the vicinity of HIV-1 budding sites by cryo-electron tomography (cET). Filamentous structures emanating from viral buds and suggested to correspond to actin filaments have been observed by atomic force microscopy. To determine whether the NC domain of Gag is required for actin association with viral buds and for actin incorporation into HIV-1, we performed comparative analyses of virus-like particles (VLPs) obtained by expression of wild-type HIV-1 Gag or a Gag variant where the entire NC domain had been replaced by a dimerizing leucine zipper [Gag(LZ)]. The latter protein yielded efficient production of VLPs with near-wild-type assembly kinetics and size and exhibited a regular immature Gag lattice. Typical HIV-1 budding sites were detected by using cET in cells expressing either Gag or Gag(LZ), and no difference was observed regarding the association of buds with the F-actin network. Furthermore, actin was equally incorporated into wild-type HIV-1 and Gag- or Gag(LZ)-derived VLPs, with less actin per particle observed than had been reported previously. Incorporation appeared to correlate with the relative intracellular actin concentration, suggesting an uptake of cytosol rather than a specific recruitment of actin. Thus, the NC domain in HIV-1 Gag does not appear to have a role in actin recruitment or actin incorporation into HIV-1 particles.

Original languageEnglish
Pages (from-to)7893-7903
Number of pages11
JournalJournal of Virology
Volume88
Issue number14
DOIs
Publication statusPublished - Jul 2014
Externally publishedYes

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