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Proteasome inhibitors enhance bacteriophage lambda (λ) mediated gene transfer in mammalian cells

Bacteriophage lambda vectors can transfer their genomes into mammalian cells, resulting in expression of phage-encoded genes. However, this process is inefficient. Experiments were therefore conducted to delineate the rate limiting step(s) involved, using a phage vector that contains a mammalian luc...

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Detalles Bibliográficos
Autores principales: Volcy, Ketna, Dewhurst, Stephen
Formato: Texto
Lenguaje:English
Publicado: Elsevier Inc. 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2654414/
https://www.ncbi.nlm.nih.gov/pubmed/19064273
http://dx.doi.org/10.1016/j.virol.2008.11.019
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author Volcy, Ketna
Dewhurst, Stephen
author_facet Volcy, Ketna
Dewhurst, Stephen
author_sort Volcy, Ketna
collection PubMed
description Bacteriophage lambda vectors can transfer their genomes into mammalian cells, resulting in expression of phage-encoded genes. However, this process is inefficient. Experiments were therefore conducted to delineate the rate limiting step(s) involved, using a phage vector that contains a mammalian luciferase reporter gene cassette. The efficiency of phage-mediated gene transfer in mammalian cells was quantitated, in the presence or absence of pharmacologic inhibitors of cell uptake and degradation pathways. Inhibitors of lysosomal proteases and proteasome inhibitors strongly enhanced phage-mediated luciferase expression, suggesting that these pathways contribute to the destruction of intracellular phage particles. In contrast, inhibition of endosome acidification had no effect on phage-mediated gene transfer, presumably because phage lambda is tolerant to extended exposure to low pH. These findings provide insights into the pathways by which phage vectors enter and transduce mammalian cells, and suggest that it may be possible to pharmacologically enhance the efficiency of phage-mediated gene transfer in mammalian cells. Finally, the data also suggest that the proteasome complex may serve as an innate defense mechanism that restricts the infection of mammalian cells by diverse viral agents.
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spelling pubmed-26544142010-02-05 Proteasome inhibitors enhance bacteriophage lambda (λ) mediated gene transfer in mammalian cells Volcy, Ketna Dewhurst, Stephen Virology Article Bacteriophage lambda vectors can transfer their genomes into mammalian cells, resulting in expression of phage-encoded genes. However, this process is inefficient. Experiments were therefore conducted to delineate the rate limiting step(s) involved, using a phage vector that contains a mammalian luciferase reporter gene cassette. The efficiency of phage-mediated gene transfer in mammalian cells was quantitated, in the presence or absence of pharmacologic inhibitors of cell uptake and degradation pathways. Inhibitors of lysosomal proteases and proteasome inhibitors strongly enhanced phage-mediated luciferase expression, suggesting that these pathways contribute to the destruction of intracellular phage particles. In contrast, inhibition of endosome acidification had no effect on phage-mediated gene transfer, presumably because phage lambda is tolerant to extended exposure to low pH. These findings provide insights into the pathways by which phage vectors enter and transduce mammalian cells, and suggest that it may be possible to pharmacologically enhance the efficiency of phage-mediated gene transfer in mammalian cells. Finally, the data also suggest that the proteasome complex may serve as an innate defense mechanism that restricts the infection of mammalian cells by diverse viral agents. Elsevier Inc. 2009-02-05 2008-12-06 /pmc/articles/PMC2654414/ /pubmed/19064273 http://dx.doi.org/10.1016/j.virol.2008.11.019 Text en Copyright © 2008 Elsevier Inc. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Article
Volcy, Ketna
Dewhurst, Stephen
Proteasome inhibitors enhance bacteriophage lambda (λ) mediated gene transfer in mammalian cells
title Proteasome inhibitors enhance bacteriophage lambda (λ) mediated gene transfer in mammalian cells
title_full Proteasome inhibitors enhance bacteriophage lambda (λ) mediated gene transfer in mammalian cells
title_fullStr Proteasome inhibitors enhance bacteriophage lambda (λ) mediated gene transfer in mammalian cells
title_full_unstemmed Proteasome inhibitors enhance bacteriophage lambda (λ) mediated gene transfer in mammalian cells
title_short Proteasome inhibitors enhance bacteriophage lambda (λ) mediated gene transfer in mammalian cells
title_sort proteasome inhibitors enhance bacteriophage lambda (λ) mediated gene transfer in mammalian cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2654414/
https://www.ncbi.nlm.nih.gov/pubmed/19064273
http://dx.doi.org/10.1016/j.virol.2008.11.019
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