Comparative analysis of virus-host interactomes with a mammalian high-throughput protein complementation assay based on Gaussia princeps luciferase.
Comparative interactomics is a strategy for inferring potential interactions among orthologous proteins or "interologs". Herein we focus, in contrast to standard homology-based inference, on the divergence of protein interaction profiles among closely related organisms, showing that the approach can correlate specific traits to phenotypic differences. As a model, this new comparative interactomic approach was applied at a large scale to human papillomaviruses (HPVs) proteins. The oncogenic potential of HPVs is mainly determined by the E6 and E7 early proteins. We have mapped and overlapped the virus-host protein interaction networks of E6 and E7 proteins from 11 distinct HPV genotypes, selected for their different tropisms and pathologies. We generated robust and comprehensive datasets by combining two orthogonal protein interaction assays: yeast two-hybrid (Y2H), and our recently described "high-throughput Gaussia princeps protein complementation assay" (HT-GPCA). HT-GPCA detects protein interaction by measuring the interaction-mediated reconstitution of activity of a split G. princeps luciferase. Hierarchical clustering of interaction profiles recapitulated HPV phylogeny and was used to correlate specific virus-host interaction profiles with pathological traits, reflecting the distinct carcinogenic potentials of different HPVs. This comparative interactomics constitutes a reliable and powerful strategy to decipher molecular relationships in virtually any combination of microorganism-host interactions.
Unité de Génétique, Papillomavirus et Cancer Humain (GPCH), Institut Pasteur, Paris, France.
Grégory
G
Neveu
Patricia
P
Cassonnet
Pierre-Olivier
PO
Vidalain
Caroline
C
Rolloy
José
J
Mendoza
Louis
L
Jones
Frédéric
F
Tangy
Mandy
M
Muller
Caroline
C
Demeret
Lionel
L
Tafforeau
Vincent
V
Lotteau
Chantal
C
Rabourdin-Combe
Gilles
G
Travé
Amélie
A
Dricot
David E
DE
Hill
Marc
M
Vidal
Michel
M
Favre
Yves
Y
Jacob
D052685:Alphapapillomavirus / Q000235:genetics / Q000502:physiology*; D028023:Arecaceae / Q000201:enzymology; D015415:Biomarkers / Q000378:metabolism; D016000:Cluster Analysis; D005838:Genotype; D057809:HEK293 Cells; D054884:Host-Pathogen Interactions; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics*; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000378:metabolism; D050725:Papillomavirus E7 Proteins / Q000235:genetics / Q000378:metabolism; D010802:Phylogeny; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics*; D011485:Protein Binding; D025941:Protein Interaction Mapping; D060066:Protein Interaction Maps; D020543:Proteome / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D020798:Two-Hybrid System Techniques; D056189:Viral Tropism
D015415:Biomarkers; C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D010940:Plant Proteins; D020543:Proteome; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D008156:Luciferases