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

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Abstract

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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Acknowledgments

This work was partially funded by a grant from the Italian Ministry of Research (MIUR; DD 1105/2002) to TC, and by a CNR Short-Term Mobility grant to NS. Assistance of Mr. G. Guarino with artwork and Mr. R. Nocerino (CNR-IBBR, Portici, Italy) with plant growth are gratefully acknowledged. We also thank Dr. S. Grillo (CNR-IBBR, Portici Italy) and Dr. A. Maggio (University of Naples ‘Federico II’, Italy) for the critical review of the manuscript. HIV-1 antisera (specific to the p24 and p17 subunits) were provided by the NIBSC Centralized Facility for AIDS reagent (EU Programme EVA, UK Medical Research Council).

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Authors and Affiliations

  1. CNR-IBBR, Institute of Biosciences and BioResources, National Research Council of Italy, Via Università 133, 80055, Portici, NA, Italy

    N. Scotti, L. Sannino & T. Cardi

  2. Max Planck Institute of Molecular Plant Physiology, Am Mühlenberg 1, 14476, Potsdam-Golm, Germany

    A. Idoine & R. Bock

  3. CNRS-IBMC, Institut de Biologie Moléculaire et Cellulaire, Centre National de la Recherche Scientifique, Plateforme Protéomique Strasbourg – Esplanade, 15 Rue Descartes, 67084, Strasbourg Cedex, France

    P. Hamman

  4. CNR-IVV, Institute of Plant Virology, National Research Council of Italy, Via G. Amendola 165/A, 70126, Bari, Italy

    A. De Stradis

  5. CNR-ISAFOM, Institute for Agricultural and Forestry Systems in the Mediterranean, National Research Council of Italy, Via Patacca 85, 80056, Ercolano, NA, Italy

    P. Giorio

  6. CNRS-IBMP, Institut de Biologie Moléculaire des Plantes, Centre National de la Recherche Scientifique, 12, rue du général Zimmer, 67084, Strasbourg Cedex, France

    L. Maréchal-Drouard

  7. Consiglio per la Ricerca e la Sperimentazione in Agricoltura – Centro di Ricerca per l’Orticoltura (CRA-ORT), Via dei Cavalleggeri 25, 84098, Pontecagnano, SA, Italy

    T. Cardi

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  1. N. Scotti
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Correspondence to N. Scotti.

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Scotti, N., Sannino, L., Idoine, A. et al. 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 Res 24, 319–331 (2015). https://doi.org/10.1007/s11248-014-9845-5

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