Isolation of suppressor genes that restore retrovirus susceptibility to a virus-resistant cell line
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BioMed Central
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Isolation of suppressor genes that restore retrovirus susceptibility to a virus-resistant cell line Guangxia Gao1,2 and Stephen P Goff*1 Address: 1Department of Biochemistry and Molecular Biophysics Howard Hughes Medical Institute Columbia University College of Physicians and Surgeons New York NY 10032, USA and 2Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China Email: Guangxia Gao - [email protected]; Stephen P Goff* - [email protected] * Corresponding author
Published: 28 September 2004 Retrovirology 2004, 1:30
doi:10.1186/1742-4690-1-30
Received: 24 August 2004 Accepted: 28 September 2004
This article is available from: http://www.retrovirology.com/content/1/1/30 © 2004 Gao and Goff; licensee BioMed Central Ltd. This is an open-access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract Background: Genetic selections in mammalian cell lines have recently been developed for the isolation of mutant cells that are refractory to infection by retroviruses. These selections have been used to recover lines that block early postentry stages of infection, either before reverse transcription or before nuclear entry. The mechanisms of action of these blocks remain unknown. Results: We have devised a method for the selection of genes from cDNA libraries that suppress the block to virus infection, and so restore virus susceptibility. The protocol involves the transformation of pools of resistant cells by cDNA expression libraries, followed by the selection for rare virus-sensitive cells, using multiple rounds of selection after infection by marked viral vector genomes. The suppressor genes were then recovered from these virus sensitive cells, and their ability to restore virus susceptibility was confirmed by reintroduction of these cDNAs into the resistant line. Conclusions: The identities of these genes provide insights into the mechanism of virus resistance and will help to define new pathways used during retrovirus infection. The methods for gene isolation developed here will also permit the identification of similar suppressors that modify or override other recently identified virus resistance genes.
Background It is becoming increasingly apparent that mammalian cells harbor numerous genes that induce intracellular blocks to retrovirus infection [1,2]. These genes have presumably evolved and been maintained in the genome in response to the pathogenic and lethal consequences of infection, and are now thought to constitute an important part of the host defense against these viruses. Some of the genes and gene products responsible for this resistance have been recently identified, including the Fv1 locus in the mouse, which blocks infection after reverse transcription but before nuclear entry and establishment of the
integrated provirus
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