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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...
Autores principales: | , |
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Formato: | Texto |
Lenguaje: | English |
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Elsevier Inc.
2009
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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. |
format | Text |
id | pubmed-2654414 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2009 |
publisher | Elsevier Inc. |
record_format | MEDLINE/PubMed |
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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