Host Cell S Phase Restricts <italic toggle="yes">Legionella pneumophila</italic> Intracellular Replication by Destabilizing the Membrane-Bound Replication Compartment

ABSTRACT Legionella pneumophila grows within cells ranging from environmental amoebae to human macrophages. In spite of this conserved strategy of pathogenesis, identification of host factors that restrict L. pneumophila intracellular replication has not been extended outside components of the mamma...

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Autores principales:	Dennise A. de Jesús-Díaz, Connor Murphy, Asaf Sol, Marion Dorer, Ralph R. Isberg
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2017
Materias:	Drosophila Legionella pneumophila RNA interference cell cycle intracellular bacteria secretion systems Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/d9d2a1edb61a4e31965d77f1281aca4e
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spelling	oai:doaj.org-article:d9d2a1edb61a4e31965d77f1281aca4e2021-11-15T15:51:43ZHost Cell S Phase Restricts <italic toggle="yes">Legionella pneumophila</italic> Intracellular Replication by Destabilizing the Membrane-Bound Replication Compartment10.1128/mBio.02345-162150-7511https://doaj.org/article/d9d2a1edb61a4e31965d77f1281aca4e2017-09-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02345-16https://doaj.org/toc/2150-7511ABSTRACT Legionella pneumophila grows within cells ranging from environmental amoebae to human macrophages. In spite of this conserved strategy of pathogenesis, identification of host factors that restrict L. pneumophila intracellular replication has not been extended outside components of the mammalian innate immune response. We performed a double-stranded RNA (dsRNA) screen against more than 50% of the Drosophila melanogaster annotated open reading frames (ORFs) to identify host cell factors that restrict L. pneumophila. The majority of analyzed dsRNAs that stimulated L. pneumophila intracellular replication were directed against host proteins involved in protein synthesis or cell cycle control. Consistent with disruption of the cell cycle stimulating intracellular replication, proteins involved in translation initiation also resulted in G1 arrest. Stimulation of replication was dependent on the stage of cell cycle arrest, as dsRNAs causing arrest during S phase had an inhibitory effect on intracellular replication. The inhibitory effects of S phase arrest could be recapitulated in a human cell line, indicating that cell cycle control of L. pneumophila replication is evolutionarily conserved. Synchronized HeLa cell populations in S phase and challenged with L. pneumophila failed to progress through the cell cycle and were depressed for supporting intracellular replication. Poor bacterial replication in S phase was associated with loss of the vacuole membrane barrier, resulting in exposure of bacteria to the cytosol and their eventual degradation. These results are consistent with the model that S phase is inhibitory for L. pneumophila intracellular survival as a consequence of failure to maintain the integrity of the membrane surrounding intracellular bacteria. IMPORTANCE Legionella pneumophila has the ability to replicate within human macrophages and amoebal hosts. Here, we report that the host cell cycle influences L. pneumophila intracellular replication. Our data demonstrate that the G1 and G2/M phases of the host cell cycle are permissive for bacterial replication, while S phase is toxic for the bacterium. L. pneumophila replicates poorly within host cells present in S phase. The inability of L. pneumophila to replicate relies on its failure to control the integrity of its vacuole, leading to cytosolic exposure of the bacteria and eventual degradation. The data presented here argue that growth-arrested host cells that are encountered by L. pneumophila in either the environment or within human hosts are ideal targets for intracellular replication because their transit through S phase is blocked.Dennise A. de Jesús-DíazConnor MurphyAsaf SolMarion DorerRalph R. IsbergAmerican Society for MicrobiologyarticleDrosophilaLegionella pneumophilaRNA interferencecell cycleintracellular bacteriasecretion systemsMicrobiologyQR1-502ENmBio, Vol 8, Iss 4 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Drosophila Legionella pneumophila RNA interference cell cycle intracellular bacteria secretion systems Microbiology QR1-502
spellingShingle	Drosophila Legionella pneumophila RNA interference cell cycle intracellular bacteria secretion systems Microbiology QR1-502 Dennise A. de Jesús-Díaz Connor Murphy Asaf Sol Marion Dorer Ralph R. Isberg Host Cell S Phase Restricts <italic toggle="yes">Legionella pneumophila</italic> Intracellular Replication by Destabilizing the Membrane-Bound Replication Compartment
description	ABSTRACT Legionella pneumophila grows within cells ranging from environmental amoebae to human macrophages. In spite of this conserved strategy of pathogenesis, identification of host factors that restrict L. pneumophila intracellular replication has not been extended outside components of the mammalian innate immune response. We performed a double-stranded RNA (dsRNA) screen against more than 50% of the Drosophila melanogaster annotated open reading frames (ORFs) to identify host cell factors that restrict L. pneumophila. The majority of analyzed dsRNAs that stimulated L. pneumophila intracellular replication were directed against host proteins involved in protein synthesis or cell cycle control. Consistent with disruption of the cell cycle stimulating intracellular replication, proteins involved in translation initiation also resulted in G1 arrest. Stimulation of replication was dependent on the stage of cell cycle arrest, as dsRNAs causing arrest during S phase had an inhibitory effect on intracellular replication. The inhibitory effects of S phase arrest could be recapitulated in a human cell line, indicating that cell cycle control of L. pneumophila replication is evolutionarily conserved. Synchronized HeLa cell populations in S phase and challenged with L. pneumophila failed to progress through the cell cycle and were depressed for supporting intracellular replication. Poor bacterial replication in S phase was associated with loss of the vacuole membrane barrier, resulting in exposure of bacteria to the cytosol and their eventual degradation. These results are consistent with the model that S phase is inhibitory for L. pneumophila intracellular survival as a consequence of failure to maintain the integrity of the membrane surrounding intracellular bacteria. IMPORTANCE Legionella pneumophila has the ability to replicate within human macrophages and amoebal hosts. Here, we report that the host cell cycle influences L. pneumophila intracellular replication. Our data demonstrate that the G1 and G2/M phases of the host cell cycle are permissive for bacterial replication, while S phase is toxic for the bacterium. L. pneumophila replicates poorly within host cells present in S phase. The inability of L. pneumophila to replicate relies on its failure to control the integrity of its vacuole, leading to cytosolic exposure of the bacteria and eventual degradation. The data presented here argue that growth-arrested host cells that are encountered by L. pneumophila in either the environment or within human hosts are ideal targets for intracellular replication because their transit through S phase is blocked.
format	article
author	Dennise A. de Jesús-Díaz Connor Murphy Asaf Sol Marion Dorer Ralph R. Isberg
author_facet	Dennise A. de Jesús-Díaz Connor Murphy Asaf Sol Marion Dorer Ralph R. Isberg
author_sort	Dennise A. de Jesús-Díaz
title	Host Cell S Phase Restricts <italic toggle="yes">Legionella pneumophila</italic> Intracellular Replication by Destabilizing the Membrane-Bound Replication Compartment
title_short	Host Cell S Phase Restricts <italic toggle="yes">Legionella pneumophila</italic> Intracellular Replication by Destabilizing the Membrane-Bound Replication Compartment
title_full	Host Cell S Phase Restricts <italic toggle="yes">Legionella pneumophila</italic> Intracellular Replication by Destabilizing the Membrane-Bound Replication Compartment
title_fullStr	Host Cell S Phase Restricts <italic toggle="yes">Legionella pneumophila</italic> Intracellular Replication by Destabilizing the Membrane-Bound Replication Compartment
title_full_unstemmed	Host Cell S Phase Restricts <italic toggle="yes">Legionella pneumophila</italic> Intracellular Replication by Destabilizing the Membrane-Bound Replication Compartment
title_sort	host cell s phase restricts <italic toggle="yes">legionella pneumophila</italic> intracellular replication by destabilizing the membrane-bound replication compartment
publisher	American Society for Microbiology
publishDate	2017
url	https://doaj.org/article/d9d2a1edb61a4e31965d77f1281aca4e
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Host Cell S Phase Restricts <italic toggle="yes">Legionella pneumophila</italic> Intracellular Replication by Destabilizing the Membrane-Bound Replication Compartment

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