Disclosing bacillus virus BAM35 and its host. Identification and characterization of the viral SSB, host genomic characterization and phage-bacteria interactome

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 15-12-2020

Detalhes bibliográficos
Autor: Lechuga Mateo, Ana
Formato: tesis doctoral
Fecha de publicación:2020
País:España
Recursos:Universidad Autónoma de Madrid
Repositorio:Biblos-e Archivo. Repositorio Institucional de la UAM
Idioma:inglés
OAI Identifier:oai:repositorio.uam.es:10486/696299
Acesso em linha:http://hdl.handle.net/10486/696299
Access Level:acceso abierto
Palavra-chave:Bacterias gram-positivas
Virología
Bacteriófagos
Biología y Biomedicina / Biología
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network_name_str España
repository_id_str
dc.title.none.fl_str_mv Disclosing bacillus virus BAM35 and its host. Identification and characterization of the viral SSB, host genomic characterization and phage-bacteria interactome
Estudio de Bacillus virus Bam35 y su hospedador. Identificación y caracterización de la SSB viral, caracterización genómica del hospedador e interactoma fago-bacteria
title Disclosing bacillus virus BAM35 and its host. Identification and characterization of the viral SSB, host genomic characterization and phage-bacteria interactome
spellingShingle Disclosing bacillus virus BAM35 and its host. Identification and characterization of the viral SSB, host genomic characterization and phage-bacteria interactome
Lechuga Mateo, Ana
Bacterias gram-positivas
Virología
Bacteriófagos
Biología y Biomedicina / Biología
title_short Disclosing bacillus virus BAM35 and its host. Identification and characterization of the viral SSB, host genomic characterization and phage-bacteria interactome
title_full Disclosing bacillus virus BAM35 and its host. Identification and characterization of the viral SSB, host genomic characterization and phage-bacteria interactome
title_fullStr Disclosing bacillus virus BAM35 and its host. Identification and characterization of the viral SSB, host genomic characterization and phage-bacteria interactome
title_full_unstemmed Disclosing bacillus virus BAM35 and its host. Identification and characterization of the viral SSB, host genomic characterization and phage-bacteria interactome
title_sort Disclosing bacillus virus BAM35 and its host. Identification and characterization of the viral SSB, host genomic characterization and phage-bacteria interactome
dc.creator.none.fl_str_mv Lechuga Mateo, Ana
author Lechuga Mateo, Ana
author_facet Lechuga Mateo, Ana
author_role author
dc.contributor.none.fl_str_mv Salas Falgueras, Margarita
Redrejo Rodríguez, Modesto
Departamento de Biología Molecular
Facultad de Ciencias
UAM-CSIC. Centro de Biología Molecular Severo Ochoa (CBM)
dc.subject.none.fl_str_mv Bacterias gram-positivas
Virología
Bacteriófagos
Biología y Biomedicina / Biología
topic Bacterias gram-positivas
Virología
Bacteriófagos
Biología y Biomedicina / Biología
description Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 15-12-2020
publishDate 2020
dc.date.none.fl_str_mv 2020
2020-12-15
dc.type.none.fl_str_mv doctoral thesis
http://purl.org/coar/resource_type/c_db06
NA
http://purl.org/coar/version/c_be7fb7dd8ff6fe43
dc.type.openaire.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
dc.identifier.none.fl_str_mv http://hdl.handle.net/10486/696299
url http://hdl.handle.net/10486/696299
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Biblos-e Archivo. Repositorio Institucional de la UAM
instname:Universidad Autónoma de Madrid
instname_str Universidad Autónoma de Madrid
reponame_str Biblos-e Archivo. Repositorio Institucional de la UAM
collection Biblos-e Archivo. Repositorio Institucional de la UAM
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869418007083614208
spelling Disclosing bacillus virus BAM35 and its host. Identification and characterization of the viral SSB, host genomic characterization and phage-bacteria interactomeEstudio de Bacillus virus Bam35 y su hospedador. Identificación y caracterización de la SSB viral, caracterización genómica del hospedador e interactoma fago-bacteriaLechuga Mateo, AnaBacterias gram-positivasVirologíaBacteriófagosBiología y Biomedicina / BiologíaTesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 15-12-2020Esta tesis tiene embargado el acceso al texto completo hasta el 15-06-2022Bacillus virus Bam35 is the main model virus of the Betatectivirus genus, which belongs to the Tectiviridae family of tailless, icosahedral, emembrane-containing bacteriophages. The interest in these viruses has greatly increased in recent years, because of its possible evolutionary relationship with various groups of prokaryotic and eukaryotic viruses. Also, betatectiviruses infect Bacillus cereus s. l. members, known by their pathogenicity and use in industry. Here, we address Bam35 biology from a broad perspective, from the particular characterization of its single-stranded DNA-binding protein (SSB) to the characterization of its host and the protein-protein interactions (PPIs) with it. The protein-primed DNA replication mechanism of Bam35 has been recently elucidated although no accessory proteins in this process have been described to date. In this work, we have identified and characterized the SSB of Bam35, B35SSB. We described B35SSB as a member of a novel group of SSBs with a conserved fold, highly divergent from the canonical OB-fold. This group mainly includes SSBs from distantly related phages with protein-primed replication. The biochemical characterization of B35SSB showed a high capacity to bind ssDNA although binding to dsDNA and RNA seems to be negligible. B35SSB is a monomer in solution that multimerizes along the substrate in the presence of ssDNA. Thus, B35SSB binds ssDNA in a non-distributive and sequence-independent manner with very high cooperativity, resulting in a high binding efficiency. We have also identified several aromatic and positively charged conserved residues essential for ssDNA-binding capacity, being aromatic residues essential for protein-DNA interaction. Moreover, B35SSB stimulates in vitro processive DNA synthesis, increasing the reaction yield without specificity for its cognate DNA polymerase. Several studies addressing Bam35 features have been performed. However, the Bacillus strains commonly used to study this phage have not been characterized. Thus, we have obtained the complete genome of B. thuringiensis HER1410, a known host strain for a variety of Bacillus phages and particularly useful for Bam35 studies. We have fully resolved the genome enabling the delineation of three plasmids: pLUSID1, a likely conjugative plasmid involved in virulence; pLUSID2, potentially related to sporulation, and a smaller plasmidial prophage pLUSID3, with a dual lifestyle whose integration within the chromosome causes the disruption of a key flagellar gene. We also detected a unique location of the single cry gene, cry1Ba4, found on a genomic island near the chromosomal replication origin. Phylogenetic analysis placed this strain next to B. thuringiensis sv. entomocidus and a within a clade comprising different B. thuringiensis and with B. cereus s. s. strains, in agreement with the intermingled taxonomy of the B. cereus group. Finally, we present the first wide analysis of protein-protein interactions for the Bam35-B. thuringiensis virushost model using a novel approach in which we combined the traditional yeast two-hybrid (Y2H) system and Illumina high-throughput sequencing technology. We generated and thoroughly analyzed a genomic library of Bam35’s host B. thuringiensis HER1410 and used it to screen interactions with all the viral proteins. A total of 158 screens have resulted in the detection of more than 4,000 putative interactions from which we obtained 183 highly-selective interactions that define a proposed virus-host interactome. Overall, host metabolism proteins and peptidases are particularly enriched in the interactome, showing differing patterns to the known host-phages PPIs. Besides, we have also detected specific and novel host-viral PPIs which have allowed us to propose functions for several Bam35 unknown function proteins and achieve a better understanding of the Bam35-B. thuringiensis interactionSalas Falgueras, MargaritaRedrejo Rodríguez, ModestoDepartamento de Biología MolecularFacultad de CienciasUAM-CSIC. Centro de Biología Molecular Severo Ochoa (CBM)20202020-12-15doctoral thesishttp://purl.org/coar/resource_type/c_db06NAhttp://purl.org/coar/version/c_be7fb7dd8ff6fe43info:eu-repo/semantics/doctoralThesisapplication/pdfhttp://hdl.handle.net/10486/696299reponame:Biblos-e Archivo. Repositorio Institucional de la UAMinstname:Universidad Autónoma de MadridInglésengopen accesshttp://purl.org/coar/access_right/c_abf2info:eu-repo/semantics/openAccessoai:repositorio.uam.es:10486/6962992026-06-23T12:46:27Z
score 15,300719

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