A Functional Role for ADAM10 in Human Immunodeficiency Virus Type-1 Replication

<p>Abstract</p> <p>Background</p> <p>Gene trap insertional mutagenesis was used as a high-throughput approach to discover cellular genes participating in viral infection by screening libraries of cells selected for survival from lytic infection with a variety of viruses...

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Main Authors: Rubin Donald H, Hodge Thomas W, Sheng Jinsong, Li Guangyu, Murray James L, Friedrich Brian M, O'Brien William A, Ferguson Monique R
Format: Article
Language:English
Published: BMC 2011-05-01
Series:Retrovirology
Online Access:http://www.retrovirology.com/content/8/1/32
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spelling doaj-7a1921e0b470485aad81c151b2156eb92020-11-25T00:21:44ZengBMCRetrovirology1742-46902011-05-01813210.1186/1742-4690-8-32A Functional Role for ADAM10 in Human Immunodeficiency Virus Type-1 ReplicationRubin Donald HHodge Thomas WSheng JinsongLi GuangyuMurray James LFriedrich Brian MO'Brien William AFerguson Monique R<p>Abstract</p> <p>Background</p> <p>Gene trap insertional mutagenesis was used as a high-throughput approach to discover cellular genes participating in viral infection by screening libraries of cells selected for survival from lytic infection with a variety of viruses. Cells harboring a disrupted <it>ADAM10 </it>(A Disintegrin and Metalloprotease 10) allele survived reovirus infection, and subsequently ADAM10 was shown by RNA interference to be important for replication of HIV-1.</p> <p>Results</p> <p>Silencing ADAM10 expression with small interfering RNA (siRNA) 48 hours before infection significantly inhibited HIV-1 replication in primary human monocyte-derived macrophages and in CD4<sup>+ </sup>cell lines. In agreement, ADAM10 over-expression significantly increased HIV-1 replication. ADAM10 down-regulation did not inhibit viral reverse transcription, indicating that viral entry and uncoating are also independent of ADAM10 expression. Integration of HIV-1 cDNA was reduced in ADAM10 down-regulated cells; however, concomitant 2-LTR circle formation was not detected, suggesting that HIV-1 does not enter the nucleus. Further, ADAM10 silencing inhibited downstream reporter gene expression and viral protein translation. Interestingly, we found that while the metalloprotease domain of ADAM10 is not required for HIV-1 replication, ADAM15 and γ-secretase (which proteolytically release the extracellular and intracellular domains of ADAM10 from the plasma membrane, respectively) do support productive infection.</p> <p>Conclusions</p> <p>We propose that ADAM10 facilitates replication at the level of nuclear trafficking. Collectively, our data support a model whereby ADAM10 is cleaved by ADAM15 and γ-secretase and that the ADAM10 intracellular domain directly facilitates HIV-1 nuclear trafficking. Thus, ADAM10 represents a novel cellular target class for development of antiretroviral drugs.</p> http://www.retrovirology.com/content/8/1/32
collection DOAJ
language English
format Article
sources DOAJ
author Rubin Donald H
Hodge Thomas W
Sheng Jinsong
Li Guangyu
Murray James L
Friedrich Brian M
O'Brien William A
Ferguson Monique R
spellingShingle Rubin Donald H
Hodge Thomas W
Sheng Jinsong
Li Guangyu
Murray James L
Friedrich Brian M
O'Brien William A
Ferguson Monique R
A Functional Role for ADAM10 in Human Immunodeficiency Virus Type-1 Replication
Retrovirology
author_facet Rubin Donald H
Hodge Thomas W
Sheng Jinsong
Li Guangyu
Murray James L
Friedrich Brian M
O'Brien William A
Ferguson Monique R
author_sort Rubin Donald H
title A Functional Role for ADAM10 in Human Immunodeficiency Virus Type-1 Replication
title_short A Functional Role for ADAM10 in Human Immunodeficiency Virus Type-1 Replication
title_full A Functional Role for ADAM10 in Human Immunodeficiency Virus Type-1 Replication
title_fullStr A Functional Role for ADAM10 in Human Immunodeficiency Virus Type-1 Replication
title_full_unstemmed A Functional Role for ADAM10 in Human Immunodeficiency Virus Type-1 Replication
title_sort functional role for adam10 in human immunodeficiency virus type-1 replication
publisher BMC
series Retrovirology
issn 1742-4690
publishDate 2011-05-01
description <p>Abstract</p> <p>Background</p> <p>Gene trap insertional mutagenesis was used as a high-throughput approach to discover cellular genes participating in viral infection by screening libraries of cells selected for survival from lytic infection with a variety of viruses. Cells harboring a disrupted <it>ADAM10 </it>(A Disintegrin and Metalloprotease 10) allele survived reovirus infection, and subsequently ADAM10 was shown by RNA interference to be important for replication of HIV-1.</p> <p>Results</p> <p>Silencing ADAM10 expression with small interfering RNA (siRNA) 48 hours before infection significantly inhibited HIV-1 replication in primary human monocyte-derived macrophages and in CD4<sup>+ </sup>cell lines. In agreement, ADAM10 over-expression significantly increased HIV-1 replication. ADAM10 down-regulation did not inhibit viral reverse transcription, indicating that viral entry and uncoating are also independent of ADAM10 expression. Integration of HIV-1 cDNA was reduced in ADAM10 down-regulated cells; however, concomitant 2-LTR circle formation was not detected, suggesting that HIV-1 does not enter the nucleus. Further, ADAM10 silencing inhibited downstream reporter gene expression and viral protein translation. Interestingly, we found that while the metalloprotease domain of ADAM10 is not required for HIV-1 replication, ADAM15 and γ-secretase (which proteolytically release the extracellular and intracellular domains of ADAM10 from the plasma membrane, respectively) do support productive infection.</p> <p>Conclusions</p> <p>We propose that ADAM10 facilitates replication at the level of nuclear trafficking. Collectively, our data support a model whereby ADAM10 is cleaved by ADAM15 and γ-secretase and that the ADAM10 intracellular domain directly facilitates HIV-1 nuclear trafficking. Thus, ADAM10 represents a novel cellular target class for development of antiretroviral drugs.</p>
url http://www.retrovirology.com/content/8/1/32
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