Binding of transcription factor activating protein 2 γ on the 5′-proximal promoter region of human porcine endogenous retrovirus subgroup A receptor 2/GPR172B
Nakaya Y, Shimode S, Kobayashi T, Imakawa K, Miyazawa T. Binding of transcription factor activating protein 2γ on the 5′‐proximal promoter region of human porcine endogenous retrovirus subgroup A receptor 2/GPR172B. Xenotransplantation 2012; 19: 177–185. © 2012 John Wiley & Sons A/S. : Backgroun...
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description | Nakaya Y, Shimode S, Kobayashi T, Imakawa K, Miyazawa T. Binding of transcription factor activating protein 2γ on the 5′‐proximal promoter region of human porcine endogenous retrovirus subgroup A receptor 2/GPR172B. Xenotransplantation 2012; 19: 177–185. © 2012 John Wiley & Sons A/S.
: Background: Xenotransplantation is one of the solutions for the shortage of organ donors, and pigs have been considered to be the most suitable animal donors. Specific pathogen‐free pigs are utilized in the xenotransplantation; however, pigs have infectious gammaretroviruses, named porcine endogenous retroviruses (PERVs) in their genome. Of them, PERV‐A and PERV‐B can infect human cells in vitro and potentially induce diseases like other gammaretroviruses. The human cellular receptors for PERV‐A were identified and named human PERV‐A receptor (HuPAR)‐1 and HuPAR‐2 (also called as GPR172A and GPR172B, respectively). We have recently reported that HuPAR‐2 expression was regulated by epigenetic modification and preferentially expressed in placenta. However, the detailed mechanisms of HuPAR‐2 expression have not been fully characterized. In this study, we analyzed molecular mechanisms associated with HuPAR‐2 transcription through the identification of transcription factors that bind to the promoter region of HuPAR‐2.
Methods: In situ hybridization was performed to identify the cells expressing HuPAR‐2 in placental tissues. Transcriptional activities were measured by dual‐luciferase reporter assay using serial deletion mutants of HuPAR‐2 5′‐flanking region. To identify the transcription factors bound to the promoter region, in silico analysis, electrophoresis mobility shift assay, and chromatin immunoprecipitation assay were conducted. The effect of the transcription factor transcription factor activator protein (TFAP)‐2γ on the promoter activities was investigated by overexpression of the factor.
Results: We identified that HuPAR‐2 was specifically expressed in villous trophoblast cells. We also identified that a region spanning from −126 to −32 had proximal promoter activities and TFAP‐2γ bound to a region spanning from −58 to −35 in vitro and in vivo. The overexpression of TFAP‐2γ also augmented the proximal promoter activity.
Conclusion: We demonstrated that TFAP‐2γ is one of the transcription factors involved in the HuPAR‐2 expression in human villous trophoblast cells. By studying transcriptional factors involved in the expression of HuPAR‐2, we may find a clue to contr |
doi_str_mv | 10.1111/j.1399-3089.2012.00701.x |
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: Background: Xenotransplantation is one of the solutions for the shortage of organ donors, and pigs have been considered to be the most suitable animal donors. Specific pathogen‐free pigs are utilized in the xenotransplantation; however, pigs have infectious gammaretroviruses, named porcine endogenous retroviruses (PERVs) in their genome. Of them, PERV‐A and PERV‐B can infect human cells in vitro and potentially induce diseases like other gammaretroviruses. The human cellular receptors for PERV‐A were identified and named human PERV‐A receptor (HuPAR)‐1 and HuPAR‐2 (also called as GPR172A and GPR172B, respectively). We have recently reported that HuPAR‐2 expression was regulated by epigenetic modification and preferentially expressed in placenta. However, the detailed mechanisms of HuPAR‐2 expression have not been fully characterized. In this study, we analyzed molecular mechanisms associated with HuPAR‐2 transcription through the identification of transcription factors that bind to the promoter region of HuPAR‐2.
Methods: In situ hybridization was performed to identify the cells expressing HuPAR‐2 in placental tissues. Transcriptional activities were measured by dual‐luciferase reporter assay using serial deletion mutants of HuPAR‐2 5′‐flanking region. To identify the transcription factors bound to the promoter region, in silico analysis, electrophoresis mobility shift assay, and chromatin immunoprecipitation assay were conducted. The effect of the transcription factor transcription factor activator protein (TFAP)‐2γ on the promoter activities was investigated by overexpression of the factor.
Results: We identified that HuPAR‐2 was specifically expressed in villous trophoblast cells. We also identified that a region spanning from −126 to −32 had proximal promoter activities and TFAP‐2γ bound to a region spanning from −58 to −35 in vitro and in vivo. The overexpression of TFAP‐2γ also augmented the proximal promoter activity.
Conclusion: We demonstrated that TFAP‐2γ is one of the transcription factors involved in the HuPAR‐2 expression in human villous trophoblast cells. By studying transcriptional factors involved in the expression of HuPAR‐2, we may find a clue to control the potential risks caused by PERV‐A infection in xenotransplantation.</description><identifier>ISSN: 0908-665X</identifier><identifier>EISSN: 1399-3089</identifier><identifier>DOI: 10.1111/j.1399-3089.2012.00701.x</identifier><identifier>PMID: 22702469</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>5' Untranslated Regions - physiology ; Activating Transcription Factor 2 - metabolism ; Animals ; Buffers ; Cell Line, Tumor ; Choriocarcinoma ; Chromatin ; chromatin immunoprecipitation assay ; Deletion mutant ; Donors ; Electrophoresis ; electrophoresis mobility shift assay ; Electrophoretic mobility ; Endogenous Retroviruses - genetics ; Endogenous Retroviruses - metabolism ; epigenetics ; Female ; Genomes ; Humans ; HuPAR-2/GPR172B ; Immunoprecipitation ; in situ hybridization ; Infection ; luciferase assay ; Membrane Transport Proteins - genetics ; Membrane Transport Proteins - metabolism ; Molecular modelling ; Mutagenesis - physiology ; Placenta ; Porcine endogenous retrovirus ; Postoperative Complications - metabolism ; Postoperative Complications - virology ; Pregnancy ; Promoter Regions, Genetic - physiology ; Promoters ; Receptors, G-Protein-Coupled - genetics ; Receptors, G-Protein-Coupled - metabolism ; Retrovirus ; RNA, Messenger - metabolism ; supershift assay ; Swine ; TFAP-2γ ; Transcription factors ; Transplantation, Heterologous - adverse effects ; Transplantation, Heterologous - physiology ; Trophoblasts ; Trophoblasts - metabolism ; Trophoblasts - virology ; Xenografts ; Zoonoses</subject><ispartof>Xenotransplantation (Københaven), 2012-05, Vol.19 (3), p.177-185</ispartof><rights>2012 John Wiley & Sons A/S</rights><rights>2012 John Wiley & Sons A/S.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4391-7fe417755baefc49f8026a68a72e791fa5d3a414009569eaf649f1cb9514b2c23</citedby><cites>FETCH-LOGICAL-c4391-7fe417755baefc49f8026a68a72e791fa5d3a414009569eaf649f1cb9514b2c23</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fj.1399-3089.2012.00701.x$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fj.1399-3089.2012.00701.x$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22702469$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Nakaya, Yuki</creatorcontrib><creatorcontrib>Shimode, Sayumi</creatorcontrib><creatorcontrib>Kobayashi, Takeshi</creatorcontrib><creatorcontrib>Imakawa, Kazuhiko</creatorcontrib><creatorcontrib>Miyazawa, Takayuki</creatorcontrib><title>Binding of transcription factor activating protein 2 γ on the 5′-proximal promoter region of human porcine endogenous retrovirus subgroup A receptor 2/GPR172B</title><title>Xenotransplantation (Københaven)</title><addtitle>Xenotransplantation</addtitle><description>Nakaya Y, Shimode S, Kobayashi T, Imakawa K, Miyazawa T. Binding of transcription factor activating protein 2γ on the 5′‐proximal promoter region of human porcine endogenous retrovirus subgroup A receptor 2/GPR172B. Xenotransplantation 2012; 19: 177–185. © 2012 John Wiley & Sons A/S.
: Background: Xenotransplantation is one of the solutions for the shortage of organ donors, and pigs have been considered to be the most suitable animal donors. Specific pathogen‐free pigs are utilized in the xenotransplantation; however, pigs have infectious gammaretroviruses, named porcine endogenous retroviruses (PERVs) in their genome. Of them, PERV‐A and PERV‐B can infect human cells in vitro and potentially induce diseases like other gammaretroviruses. The human cellular receptors for PERV‐A were identified and named human PERV‐A receptor (HuPAR)‐1 and HuPAR‐2 (also called as GPR172A and GPR172B, respectively). We have recently reported that HuPAR‐2 expression was regulated by epigenetic modification and preferentially expressed in placenta. However, the detailed mechanisms of HuPAR‐2 expression have not been fully characterized. In this study, we analyzed molecular mechanisms associated with HuPAR‐2 transcription through the identification of transcription factors that bind to the promoter region of HuPAR‐2.
Methods: In situ hybridization was performed to identify the cells expressing HuPAR‐2 in placental tissues. Transcriptional activities were measured by dual‐luciferase reporter assay using serial deletion mutants of HuPAR‐2 5′‐flanking region. To identify the transcription factors bound to the promoter region, in silico analysis, electrophoresis mobility shift assay, and chromatin immunoprecipitation assay were conducted. The effect of the transcription factor transcription factor activator protein (TFAP)‐2γ on the promoter activities was investigated by overexpression of the factor.
Results: We identified that HuPAR‐2 was specifically expressed in villous trophoblast cells. We also identified that a region spanning from −126 to −32 had proximal promoter activities and TFAP‐2γ bound to a region spanning from −58 to −35 in vitro and in vivo. The overexpression of TFAP‐2γ also augmented the proximal promoter activity.
Conclusion: We demonstrated that TFAP‐2γ is one of the transcription factors involved in the HuPAR‐2 expression in human villous trophoblast cells. By studying transcriptional factors involved in the expression of HuPAR‐2, we may find a clue to control the potential risks caused by PERV‐A infection in xenotransplantation.</description><subject>5' Untranslated Regions - physiology</subject><subject>Activating Transcription Factor 2 - metabolism</subject><subject>Animals</subject><subject>Buffers</subject><subject>Cell Line, Tumor</subject><subject>Choriocarcinoma</subject><subject>Chromatin</subject><subject>chromatin immunoprecipitation assay</subject><subject>Deletion mutant</subject><subject>Donors</subject><subject>Electrophoresis</subject><subject>electrophoresis mobility shift assay</subject><subject>Electrophoretic mobility</subject><subject>Endogenous Retroviruses - genetics</subject><subject>Endogenous Retroviruses - metabolism</subject><subject>epigenetics</subject><subject>Female</subject><subject>Genomes</subject><subject>Humans</subject><subject>HuPAR-2/GPR172B</subject><subject>Immunoprecipitation</subject><subject>in situ hybridization</subject><subject>Infection</subject><subject>luciferase assay</subject><subject>Membrane Transport Proteins - genetics</subject><subject>Membrane Transport Proteins - metabolism</subject><subject>Molecular modelling</subject><subject>Mutagenesis - physiology</subject><subject>Placenta</subject><subject>Porcine endogenous retrovirus</subject><subject>Postoperative Complications - metabolism</subject><subject>Postoperative Complications - virology</subject><subject>Pregnancy</subject><subject>Promoter Regions, Genetic - physiology</subject><subject>Promoters</subject><subject>Receptors, G-Protein-Coupled - genetics</subject><subject>Receptors, G-Protein-Coupled - metabolism</subject><subject>Retrovirus</subject><subject>RNA, Messenger - metabolism</subject><subject>supershift assay</subject><subject>Swine</subject><subject>TFAP-2γ</subject><subject>Transcription factors</subject><subject>Transplantation, Heterologous - adverse effects</subject><subject>Transplantation, Heterologous - physiology</subject><subject>Trophoblasts</subject><subject>Trophoblasts - metabolism</subject><subject>Trophoblasts - virology</subject><subject>Xenografts</subject><subject>Zoonoses</subject><issn>0908-665X</issn><issn>1399-3089</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkU1u1DAUxy0EokPhCshLNkn9EcexxKYdyhSpHRAfojvL8ThTD4kd7KSd7noVrgD34BCcBIcps64X9pPf7_-e_f4AQIxynNbRJsdUiIyiSuQEYZIjxBHOt4_AbJ94DGZIoCorS3Z5AJ7FuEEIUVaxp-CAEI5IUYoZ-HFi3cq6NfQNHIJyUQfbD9Y72Cg9-ADTbq_VMCF98IOxDhL4-xdMxHBlIPtz9zNLia3tVDsRXWICDGY91UhFr8ZOOdj7oK0z0LiVXxvnx5iQIfhrG1IYx3od_NjD43SrTT_1JUeLDx8xJyfPwZNGtdG8uD8PwZe3p5_nZ9n5-8W7-fF5pgsqcMYbU2DOGauVaXQhmgqRUpWV4sRwgRvFVlQVuEBIsFIY1ZSJwboWDBc10YQegle7uukT30cTB9nZqE3bKmfSeyVGlFQiTbN6AEowJkXFaUKrHaqDjzGYRvYhjSrcJkhOXsqNnCyTk2Vy8lL-81Juk_TlfZex7sxqL_xvXgJe74Ab25rbBxeWl6fLFCR5tpPbOJjtXq7CN1lyypn8ulzI-cXZcnHxppSf6F-tjL-L</recordid><startdate>201205</startdate><enddate>201205</enddate><creator>Nakaya, Yuki</creator><creator>Shimode, Sayumi</creator><creator>Kobayashi, Takeshi</creator><creator>Imakawa, Kazuhiko</creator><creator>Miyazawa, Takayuki</creator><general>Blackwell Publishing Ltd</general><scope>BSCLL</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7U9</scope><scope>H94</scope></search><sort><creationdate>201205</creationdate><title>Binding of transcription factor activating protein 2 γ on the 5′-proximal promoter region of human porcine endogenous retrovirus subgroup A receptor 2/GPR172B</title><author>Nakaya, Yuki ; Shimode, Sayumi ; Kobayashi, Takeshi ; Imakawa, Kazuhiko ; Miyazawa, Takayuki</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4391-7fe417755baefc49f8026a68a72e791fa5d3a414009569eaf649f1cb9514b2c23</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>5' Untranslated Regions - physiology</topic><topic>Activating Transcription Factor 2 - metabolism</topic><topic>Animals</topic><topic>Buffers</topic><topic>Cell Line, Tumor</topic><topic>Choriocarcinoma</topic><topic>Chromatin</topic><topic>chromatin immunoprecipitation assay</topic><topic>Deletion mutant</topic><topic>Donors</topic><topic>Electrophoresis</topic><topic>electrophoresis mobility shift assay</topic><topic>Electrophoretic mobility</topic><topic>Endogenous Retroviruses - genetics</topic><topic>Endogenous Retroviruses - metabolism</topic><topic>epigenetics</topic><topic>Female</topic><topic>Genomes</topic><topic>Humans</topic><topic>HuPAR-2/GPR172B</topic><topic>Immunoprecipitation</topic><topic>in situ hybridization</topic><topic>Infection</topic><topic>luciferase assay</topic><topic>Membrane Transport Proteins - genetics</topic><topic>Membrane Transport Proteins - metabolism</topic><topic>Molecular modelling</topic><topic>Mutagenesis - physiology</topic><topic>Placenta</topic><topic>Porcine endogenous retrovirus</topic><topic>Postoperative Complications - metabolism</topic><topic>Postoperative Complications - virology</topic><topic>Pregnancy</topic><topic>Promoter Regions, Genetic - physiology</topic><topic>Promoters</topic><topic>Receptors, G-Protein-Coupled - genetics</topic><topic>Receptors, G-Protein-Coupled - metabolism</topic><topic>Retrovirus</topic><topic>RNA, Messenger - metabolism</topic><topic>supershift assay</topic><topic>Swine</topic><topic>TFAP-2γ</topic><topic>Transcription factors</topic><topic>Transplantation, Heterologous - adverse effects</topic><topic>Transplantation, Heterologous - physiology</topic><topic>Trophoblasts</topic><topic>Trophoblasts - metabolism</topic><topic>Trophoblasts - virology</topic><topic>Xenografts</topic><topic>Zoonoses</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Nakaya, Yuki</creatorcontrib><creatorcontrib>Shimode, Sayumi</creatorcontrib><creatorcontrib>Kobayashi, Takeshi</creatorcontrib><creatorcontrib>Imakawa, Kazuhiko</creatorcontrib><creatorcontrib>Miyazawa, Takayuki</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><jtitle>Xenotransplantation (Københaven)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Nakaya, Yuki</au><au>Shimode, Sayumi</au><au>Kobayashi, Takeshi</au><au>Imakawa, Kazuhiko</au><au>Miyazawa, Takayuki</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Binding of transcription factor activating protein 2 γ on the 5′-proximal promoter region of human porcine endogenous retrovirus subgroup A receptor 2/GPR172B</atitle><jtitle>Xenotransplantation (Københaven)</jtitle><addtitle>Xenotransplantation</addtitle><date>2012-05</date><risdate>2012</risdate><volume>19</volume><issue>3</issue><spage>177</spage><epage>185</epage><pages>177-185</pages><issn>0908-665X</issn><eissn>1399-3089</eissn><abstract>Nakaya Y, Shimode S, Kobayashi T, Imakawa K, Miyazawa T. Binding of transcription factor activating protein 2γ on the 5′‐proximal promoter region of human porcine endogenous retrovirus subgroup A receptor 2/GPR172B. Xenotransplantation 2012; 19: 177–185. © 2012 John Wiley & Sons A/S.
: Background: Xenotransplantation is one of the solutions for the shortage of organ donors, and pigs have been considered to be the most suitable animal donors. Specific pathogen‐free pigs are utilized in the xenotransplantation; however, pigs have infectious gammaretroviruses, named porcine endogenous retroviruses (PERVs) in their genome. Of them, PERV‐A and PERV‐B can infect human cells in vitro and potentially induce diseases like other gammaretroviruses. The human cellular receptors for PERV‐A were identified and named human PERV‐A receptor (HuPAR)‐1 and HuPAR‐2 (also called as GPR172A and GPR172B, respectively). We have recently reported that HuPAR‐2 expression was regulated by epigenetic modification and preferentially expressed in placenta. However, the detailed mechanisms of HuPAR‐2 expression have not been fully characterized. In this study, we analyzed molecular mechanisms associated with HuPAR‐2 transcription through the identification of transcription factors that bind to the promoter region of HuPAR‐2.
Methods: In situ hybridization was performed to identify the cells expressing HuPAR‐2 in placental tissues. Transcriptional activities were measured by dual‐luciferase reporter assay using serial deletion mutants of HuPAR‐2 5′‐flanking region. To identify the transcription factors bound to the promoter region, in silico analysis, electrophoresis mobility shift assay, and chromatin immunoprecipitation assay were conducted. The effect of the transcription factor transcription factor activator protein (TFAP)‐2γ on the promoter activities was investigated by overexpression of the factor.
Results: We identified that HuPAR‐2 was specifically expressed in villous trophoblast cells. We also identified that a region spanning from −126 to −32 had proximal promoter activities and TFAP‐2γ bound to a region spanning from −58 to −35 in vitro and in vivo. The overexpression of TFAP‐2γ also augmented the proximal promoter activity.
Conclusion: We demonstrated that TFAP‐2γ is one of the transcription factors involved in the HuPAR‐2 expression in human villous trophoblast cells. By studying transcriptional factors involved in the expression of HuPAR‐2, we may find a clue to control the potential risks caused by PERV‐A infection in xenotransplantation.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>22702469</pmid><doi>10.1111/j.1399-3089.2012.00701.x</doi><tpages>9</tpages></addata></record> |
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subjects | 5' Untranslated Regions - physiology Activating Transcription Factor 2 - metabolism Animals Buffers Cell Line, Tumor Choriocarcinoma Chromatin chromatin immunoprecipitation assay Deletion mutant Donors Electrophoresis electrophoresis mobility shift assay Electrophoretic mobility Endogenous Retroviruses - genetics Endogenous Retroviruses - metabolism epigenetics Female Genomes Humans HuPAR-2/GPR172B Immunoprecipitation in situ hybridization Infection luciferase assay Membrane Transport Proteins - genetics Membrane Transport Proteins - metabolism Molecular modelling Mutagenesis - physiology Placenta Porcine endogenous retrovirus Postoperative Complications - metabolism Postoperative Complications - virology Pregnancy Promoter Regions, Genetic - physiology Promoters Receptors, G-Protein-Coupled - genetics Receptors, G-Protein-Coupled - metabolism Retrovirus RNA, Messenger - metabolism supershift assay Swine TFAP-2γ Transcription factors Transplantation, Heterologous - adverse effects Transplantation, Heterologous - physiology Trophoblasts Trophoblasts - metabolism Trophoblasts - virology Xenografts Zoonoses |
title | Binding of transcription factor activating protein 2 γ on the 5′-proximal promoter region of human porcine endogenous retrovirus subgroup A receptor 2/GPR172B |
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