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Article

Unraveling Protein Interactions between the Temperate Virus Bam35 and Its Bacillus Host Using an Integrative Yeast Two Hybrid–High Throughput Sequencing Approach

1
Centro de Biología Molecular Severo Ochoa (CSIC-UAM), 28049 Madrid, Spain
2
Laboratory of Gene Technology, Department of Biosystems, KU Leuven, 3001 Leuven, Belgium
3
Centre of Microbial and Plant Genetics, Laboratory of Computational Systems Biology, Department of Microbial and Molecular Systems, KU Leuven, 3001 Leuven, Belgium
4
Departamento de Bioquímica, Universidad Autónoma de Madrid (UAM) and Instituto de Investigaciones Biomédicas Alberto Sols (CSIC-UAM), 28029 Madrid, Spain
*
Author to whom correspondence should be addressed.
Contributed equally to this work.
Deceased.
Academic Editor: Alicja Wegrzyn
Int. J. Mol. Sci. 2021, 22(20), 11105; https://doi.org/10.3390/ijms222011105
Received: 27 August 2021 / Revised: 8 October 2021 / Accepted: 10 October 2021 / Published: 14 October 2021
(This article belongs to the Special Issue Bacteriophage—Molecular Studies 3.0)
Bacillus virus Bam35 is the model Betatectivirus and member of the family Tectiviridae, which is composed of tailless, icosahedral, and membrane-containing bacteriophages. Interest in these viruses has greatly increased in recent years as they are thought to be an evolutionary link between diverse groups of prokaryotic and eukaryotic viruses. Additionally, betatectiviruses infect bacteria of the Bacillus cereus group, which are known for their applications in industry and notorious since it contains many pathogens. Here, we present the first protein–protein interactions (PPIs) network for a tectivirus–host system by studying the Bam35–Bacillus thuringiensis model using a novel approach that integrates the traditional yeast two-hybrid system and high-throughput sequencing (Y2H-HTS). We generated and thoroughly analyzed a genomic library of Bam35′s host B. thuringiensis HER1410 and screened interactions with all the viral proteins using different combinations of bait–prey couples. Initial analysis of the raw data enabled the identification of over 4000 candidate interactions, which were sequentially filtered 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. View Full-Text
Keywords: Bam35; tectivirus; Betatectivirus; interactome; yeast two-hybrid; Bacillus thuringiensis; protein–protein interactions; high-throughput sequencing Bam35; tectivirus; Betatectivirus; interactome; yeast two-hybrid; Bacillus thuringiensis; protein–protein interactions; high-throughput sequencing
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MDPI and ACS Style

Lechuga, A.; Lood, C.; Berjón-Otero, M.; del Prado, A.; Wagemans, J.; van Noort, V.; Lavigne, R.; Salas, M.; Redrejo-Rodríguez, M. Unraveling Protein Interactions between the Temperate Virus Bam35 and Its Bacillus Host Using an Integrative Yeast Two Hybrid–High Throughput Sequencing Approach. Int. J. Mol. Sci. 2021, 22, 11105. https://doi.org/10.3390/ijms222011105

AMA Style

Lechuga A, Lood C, Berjón-Otero M, del Prado A, Wagemans J, van Noort V, Lavigne R, Salas M, Redrejo-Rodríguez M. Unraveling Protein Interactions between the Temperate Virus Bam35 and Its Bacillus Host Using an Integrative Yeast Two Hybrid–High Throughput Sequencing Approach. International Journal of Molecular Sciences. 2021; 22(20):11105. https://doi.org/10.3390/ijms222011105

Chicago/Turabian Style

Lechuga, Ana, Cédric Lood, Mónica Berjón-Otero, Alicia del Prado, Jeroen Wagemans, Vera van Noort, Rob Lavigne, Margarita Salas, and Modesto Redrejo-Rodríguez. 2021. "Unraveling Protein Interactions between the Temperate Virus Bam35 and Its Bacillus Host Using an Integrative Yeast Two Hybrid–High Throughput Sequencing Approach" International Journal of Molecular Sciences 22, no. 20: 11105. https://doi.org/10.3390/ijms222011105

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