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Viruses 2017, 9(10), 275; doi:10.3390/v9100275

Inference of a Geminivirus−Host Protein−Protein Interaction Network through Affinity Purification and Mass Spectrometry Analysis

1
Shanghai Center for Plant Stress Biology, Chinese Academy of Sciences, Shanghai 201602, China
2
University of the Chinese Academy of Sciences, Beijing 100049, China
These authors contributed equally to this work.
*
Authors to whom correspondence should be addressed.
Received: 31 August 2017 / Revised: 20 September 2017 / Accepted: 22 September 2017 / Published: 25 September 2017
(This article belongs to the Special Issue Geminiviruses)
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Abstract

Viruses reshape the intracellular environment of their hosts, largely through protein-protein interactions, to co-opt processes necessary for viral infection and interference with antiviral defences. Due to genome size constraints and the concomitant limited coding capacity of viruses, viral proteins are generally multifunctional and have evolved to target diverse host proteins. Inference of the virus-host interaction network can be instrumental for understanding how viruses manipulate the host machinery and how re-wiring of specific pathways can contribute to disease. Here, we use affinity purification and mass spectrometry analysis (AP-MS) to define the global landscape of interactions between the geminivirus Tomato yellow leaf curl virus (TYLCV) and its host Nicotiana benthamiana. For this purpose, we expressed tagged versions of each of TYLCV-encoded proteins (C1/Rep, C2/TrAP, C3/REn, C4, V2, and CP) in planta in the presence of the virus. Using a quantitative scoring system, 728 high-confidence plant interactors were identified, and the interaction network of each viral protein was inferred; TYLCV-targeted proteins are more connected than average, and connect with other proteins through shorter paths, which would allow the virus to exert large effects with few interactions. Comparative analyses of divergence patterns between N. benthamiana and potato, a non-host Solanaceae, showed evolutionary constraints on TYLCV-targeted proteins. Our results provide a comprehensive overview of plant proteins targeted by TYLCV during the viral infection, which may contribute to uncovering the underlying molecular mechanisms of plant viral diseases and provide novel potential targets for anti-viral strategies and crop engineering. Interestingly, some of the TYLCV-interacting proteins appear to be convergently targeted by other pathogen effectors, which suggests a central role for these proteins in plant-pathogen interactions, and pinpoints them as potential targets to engineer broad-spectrum resistance to biotic stresses. View Full-Text
Keywords: Geminivirus; TYLCV; Nicotiana benthamiana; protein-protein interactions; network; hubs; broad-spectrum resistance; pathogens; plant; host Geminivirus; TYLCV; Nicotiana benthamiana; protein-protein interactions; network; hubs; broad-spectrum resistance; pathogens; plant; host
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Wang, L.; Ding, X.; Xiao, J.; Jiménez-Gόngora, T.; Liu, R.; Lozano-Durán, R. Inference of a Geminivirus−Host Protein−Protein Interaction Network through Affinity Purification and Mass Spectrometry Analysis. Viruses 2017, 9, 275.

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