The HIV-1 Pr55 gag polyprotein binds to plastidial membranes and leads to severe impairment of chloroplast biogenesis and seedling lethality in transplastomic tobacco plants.
Fiche publication
Date publication
avril 2015
Journal
Transgenic research
Auteurs
Membres identifiés du Cancéropôle Est :
Dr MARÉCHAL-DROUARD Laurence
Tous les auteurs :
Scotti N, Sannino L, Idoine A, Hamman P, De Stradis A, Giorio P, Maréchal-Drouard L, Bock R, Cardi T
Lien Pubmed
Résumé
Chloroplast genetic engineering has long been recognised as a powerful technology to produce recombinant proteins. To date, however, little attention has been given to the causes of pleiotropic effects reported, in some cases, as consequence of the expression of foreign proteins in transgenic plastids. In this study, we investigated the phenotypic alterations observed in transplastomic tobacco plants accumulating the Pr55(gag) polyprotein of human immunodeficiency virus (HIV-1). The expression of Pr55(gag) at high levels in the tobacco plastome leads to a lethal phenotype of seedlings grown in soil, severe impairment of plastid development and photosynthetic activity, with chloroplasts largely resembling undeveloped proplastids. These alterations are associated to the binding of Pr55(gag) to thylakoids. During particle assembly in HIV-1 infected human cells, the binding of Pr55(gag) to a specific lipid [phosphatidylinositol-(4-5) bisphosphate] in the plasma membrane is mediated by myristoylation at the amino-terminus and the so-called highly basic region (HBR). Surprisingly, the non-myristoylated Pr55(gag) expressed in tobacco plastids was likely able, through the HBR motif, to bind to nonphosphorous glycerogalactolipids or other classes of lipids present in plastidial membranes. Although secondary consequences of disturbed chloroplast biogenesis on expression of nuclear-encoded plastid proteins cannot be ruled out, results of proteomic analyses suggest that their altered accumulation could be due to retrograde control in which chloroplasts relay their status to the nucleus for fine-tuning of gene expression.
Mots clés
Animals, COS Cells, Cercopithecus aethiops, Chloroplasts, genetics, Fatty Acids, Monounsaturated, metabolism, HIV-1, genetics, Humans, Membranes, metabolism, Plants, Genetically Modified, genetics, Plastids, genetics, Protein Binding, Protein Precursors, genetics, Seedlings, genetics, Tobacco, genetics
Référence
Transgenic Res.. 2015 Apr;24(2):319-31