A Multifaceted Study of <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Shutdown by Virulent Podovirus LUZ19

ABSTRACT In contrast to the rapidly increasing knowledge on genome content and diversity of bacterial viruses, insights in intracellular phage development and its impact on bacterial physiology are very limited. We present a multifaceted study combining quantitative PCR (qPCR), microarray, RNA-seq,...

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Autores principales:	Rob Lavigne, Elke Lecoutere, Jeroen Wagemans, William Cenens, Abram Aertsen, Liliane Schoofs, Bart Landuyt, Jan Paeshuyse, Maurice Scheer, Max Schobert, Pieter-Jan Ceyssens
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Lenguaje:	EN
Publicado:	American Society for Microbiology 2013
Materias:	Microbiology QR1-502
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spelling	oai:doaj.org-article:bd3d43bcb42343c888404b60526e57872021-11-15T15:40:27ZA Multifaceted Study of <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Shutdown by Virulent Podovirus LUZ1910.1128/mBio.00061-132150-7511https://doaj.org/article/bd3d43bcb42343c888404b60526e57872013-05-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00061-13https://doaj.org/toc/2150-7511ABSTRACT In contrast to the rapidly increasing knowledge on genome content and diversity of bacterial viruses, insights in intracellular phage development and its impact on bacterial physiology are very limited. We present a multifaceted study combining quantitative PCR (qPCR), microarray, RNA-seq, and two-dimensional gel electrophoresis (2D-GE), to obtain a global overview of alterations in DNA, RNA, and protein content in Pseudomonas aeruginosa PAO1 cells upon infection with the strictly lytic phage LUZ19. Viral genome replication occurs in the second half of the phage infection cycle and coincides with degradation of the bacterial genome. At the RNA level, there is a sharp increase in viral mRNAs from 23 to 60% of all transcripts after 5 and 15 min of infection, respectively. Although microarray analysis revealed a complex pattern of bacterial up- and downregulated genes, the accumulation of viral mRNA clearly coincides with a general breakdown of abundant bacterial transcripts. Two-dimensional gel electrophoretic analyses shows no bacterial protein degradation during phage infection, and seven stress-related bacterial proteins appear. Moreover, the two most abundantly expressed early and late-early phage proteins, LUZ19 gene product 13 (Gp13) and Gp21, completely inhibit P. aeruginosa growth when expressed from a single-copy plasmid. Since Gp13 encodes a predicted GNAT acetyltransferase, this observation points at a crucial but yet unexplored level of posttranslational viral control during infection. IMPORTANCE Massive genome sequencing has led to important insights into the enormous genetic diversity of bacterial viruses (bacteriophages). However, for nearly all known phages, information on the impact of the phage infection on host physiology and intracellular phage development is scarce. This aspect of phage research should be revitalized, as phages evolved genes which can shut down or redirect bacterial processes in a very efficient way, which can be exploited towards antibacterial design. In this context, we initiated a study of the human opportunistic pathogen Pseudomonas aeruginosa under attack by one its most common predators, the Phikmvlikevirus. By analyzing various stages of infection at different levels, this study uncovers new features of phage infection, representing a cornerstone for future studies on members of this phage genus.Rob LavigneElke LecoutereJeroen WagemansWilliam CenensAbram AertsenLiliane SchoofsBart LanduytJan PaeshuyseMaurice ScheerMax SchobertPieter-Jan CeyssensAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 4, Iss 2 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Rob Lavigne Elke Lecoutere Jeroen Wagemans William Cenens Abram Aertsen Liliane Schoofs Bart Landuyt Jan Paeshuyse Maurice Scheer Max Schobert Pieter-Jan Ceyssens A Multifaceted Study of <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Shutdown by Virulent Podovirus LUZ19
description	ABSTRACT In contrast to the rapidly increasing knowledge on genome content and diversity of bacterial viruses, insights in intracellular phage development and its impact on bacterial physiology are very limited. We present a multifaceted study combining quantitative PCR (qPCR), microarray, RNA-seq, and two-dimensional gel electrophoresis (2D-GE), to obtain a global overview of alterations in DNA, RNA, and protein content in Pseudomonas aeruginosa PAO1 cells upon infection with the strictly lytic phage LUZ19. Viral genome replication occurs in the second half of the phage infection cycle and coincides with degradation of the bacterial genome. At the RNA level, there is a sharp increase in viral mRNAs from 23 to 60% of all transcripts after 5 and 15 min of infection, respectively. Although microarray analysis revealed a complex pattern of bacterial up- and downregulated genes, the accumulation of viral mRNA clearly coincides with a general breakdown of abundant bacterial transcripts. Two-dimensional gel electrophoretic analyses shows no bacterial protein degradation during phage infection, and seven stress-related bacterial proteins appear. Moreover, the two most abundantly expressed early and late-early phage proteins, LUZ19 gene product 13 (Gp13) and Gp21, completely inhibit P. aeruginosa growth when expressed from a single-copy plasmid. Since Gp13 encodes a predicted GNAT acetyltransferase, this observation points at a crucial but yet unexplored level of posttranslational viral control during infection. IMPORTANCE Massive genome sequencing has led to important insights into the enormous genetic diversity of bacterial viruses (bacteriophages). However, for nearly all known phages, information on the impact of the phage infection on host physiology and intracellular phage development is scarce. This aspect of phage research should be revitalized, as phages evolved genes which can shut down or redirect bacterial processes in a very efficient way, which can be exploited towards antibacterial design. In this context, we initiated a study of the human opportunistic pathogen Pseudomonas aeruginosa under attack by one its most common predators, the Phikmvlikevirus. By analyzing various stages of infection at different levels, this study uncovers new features of phage infection, representing a cornerstone for future studies on members of this phage genus.
format	article
author	Rob Lavigne Elke Lecoutere Jeroen Wagemans William Cenens Abram Aertsen Liliane Schoofs Bart Landuyt Jan Paeshuyse Maurice Scheer Max Schobert Pieter-Jan Ceyssens
author_facet	Rob Lavigne Elke Lecoutere Jeroen Wagemans William Cenens Abram Aertsen Liliane Schoofs Bart Landuyt Jan Paeshuyse Maurice Scheer Max Schobert Pieter-Jan Ceyssens
author_sort	Rob Lavigne
title	A Multifaceted Study of <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Shutdown by Virulent Podovirus LUZ19
title_short	A Multifaceted Study of <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Shutdown by Virulent Podovirus LUZ19
title_full	A Multifaceted Study of <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Shutdown by Virulent Podovirus LUZ19
title_fullStr	A Multifaceted Study of <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Shutdown by Virulent Podovirus LUZ19
title_full_unstemmed	A Multifaceted Study of <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Shutdown by Virulent Podovirus LUZ19
title_sort	multifaceted study of <named-content content-type="genus-species">pseudomonas aeruginosa</named-content> shutdown by virulent podovirus luz19
publisher	American Society for Microbiology
publishDate	2013
url	https://doaj.org/article/bd3d43bcb42343c888404b60526e5787
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A Multifaceted Study of <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Shutdown by Virulent Podovirus LUZ19

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