The HIV-1 Pr55gag polyprotein binds to plastidial membranes and leads to severe impairment of chloroplast biogenesis and seedling lethality in transplastomic tobacco plants

Transgenic Research - Tập 24 - Trang 319-331 - 2014
N. Scotti1, L. Sannino1, A. Idoine2, P. Hamman3, A. De Stradis4, P. Giorio5, L. Maréchal-Drouard6, R. Bock2, T. Cardi1,7
1CNR-IBBR, Institute of Biosciences and BioResources, National Research Council of Italy, Portici, Italy
2Max-Planck Institute of Molecular Plant Physiology, Potsdam-Golm, Germany
3CNRS-IBMC, Institut de Biologie Moléculaire et Cellulaire, Centre National de la Recherche Scientifique, Strasbourg Cedex, France
4CNR-IVV, Institute of Plant Virology, National Research Council of Italy, Bari, Italy
5CNR-ISAFOM, Institute for Agricultural and Forestry Systems in the Mediterranean, National Research Council of Italy, Ercolano, Italy
6CNRS-IBMP, Institut de Biologie Moléculaire des Plantes, Centre National de la Recherche Scientifique, Strasbourg Cedex, France
7Consiglio per la Ricerca e la Sperimentazione in Agricoltura – Centro di Ricerca per l’Orticoltura (CRA-ORT), Pontecagnano, Italy

Tóm tắt

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 Pr55gag polyprotein of human immunodeficiency virus (HIV-1). The expression of Pr55gag 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 Pr55gag to thylakoids. During particle assembly in HIV-1 infected human cells, the binding of Pr55gag 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 Pr55gag 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.

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