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Comprehensive virus-host protein interactome by the use of yeast-two-hybrid coupled to next-generation sequencing analysis

19th national competition for scientific and technical research

Intercellular Dialogue and Interactome: Pathological Implications

Senior Researcher : Margarita Salas Falgueras

Research Centre or Institution : Centro de Biología Molecular "Severo Ochoa". CSIC-Universidad Autónoma de Madrid

Abstract

The outcome of the replication cycle of a virus within the host cell depends on a large number of molecular interactions. The objective of this project is to profit from the recent development of high-throughput technologies to study virus-host interactions at the molecular level. During this year we performed a comprehensive analysis of protein-protein interactions (PPI) between the betatecvirus model Bam35 and its host Bacillus thuringiensis HER1410 coupling a yeast two-hybrid (Y2H) screening with high-throughput sequencing (HTS). Initial analysis of the raw data enabled the identification of over 4000 candidate interactions, which were sequentially filtered by technical and biological criteria to produce 182 high-confidence interactions that were defined as part of the core virus-host interactome. Overall, host metabolism proteins and peptidases were particularly enriched within the detected interactions, distinguishing this host-phage system from the other reported host-phage PPIs. Our approach also suggested biological roles for several Bam35 proteins of unknown function, including the membrane structural protein P25, which may be a viral hub with a role in host membrane modification during viral particle morphogenesis. This work resulted in a better understanding of the Bam35-B. thuringiensis interaction at the molecular level and holds great potential for the generalization of the Y2H-HTS approach for other virus-host models.

To further characterize the betatectivirus-host interactions we have recently started the study of the modification of the transcriptional landscape in Bt upon the presence of the (pro)phage under both the lysogenic and lytic cycle. Two different betatectiviruses (GIL01 and GIL16) will be analyzed upon the infection of Bacillus thuringiensis strain GBJ002, the natural host of GIL01, and after the lytic cycle induction by SOS response.

Note: Professor Modesto Redrejo Rodríguez is the main researcher of this project, since the death of Mrs. Margarita Salas Falgueras, in November 2019.