The HIV-1 Pr55^sup gag^ polyprotein binds to plastidial membranes and leads to severe impairment of chloroplast biogenesis and seedling lethality in transplastomic tobacco plants

• Published in: Transgenic research 2015-04, Vol.24 (2), p.319
• Main Authors: Scotti, N, Sannino, L, Idoine, A, Hamman, P, De Stradis, A, Giorio, P, Maréchal-drouard, L, Bock, R, Cardi, T
• Format: Article
• Language: English
• ISSN: 0962-8819; 1573-9368
• DOI: 10.1007/s11248-014-9845-5

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^sup gag^ polyprotein of human immunodeficiency virus (HIV-1). The expression of Pr55^sup 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^sup gag^ to thylakoids. During particle assembly in HIV-1 infected human cells, the binding of Pr55^sup 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^sup 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.