Retinoic Acid Receptor γ: Specific Immunodetection and Phosphorylation
Synthetic peptides corresponding to cDNA-deduced amino acid sequences unique to the human and mouse retinoic acid receptor γ1 (hRAR-γ1 and mRAR-γ1, respectively) were used to generate anti-RAR-γ1 antibodies. Four mAbs were selected, which were directed against peptides found in region A1 (Ab1γ(A1)),...
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| Veröffentlicht in: | The Journal of cell biology 1991-10, Vol.115 (2), p.535-545 |
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| container_title | The Journal of cell biology |
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| creator | Rochette-Egly, C. Lutz, Y. Saunders, M. Scheuer, I. M.-P. Gaub Chambon, P. |
| description | Synthetic peptides corresponding to cDNA-deduced amino acid sequences unique to the human and mouse retinoic acid receptor γ1 (hRAR-γ1 and mRAR-γ1, respectively) were used to generate anti-RAR-γ1 antibodies. Four mAbs were selected, which were directed against peptides found in region A1 (Ab1γ(A1)), region F (Ab2γ(mF) and Ab4γ(hF)) and region D2 (Ab5γ(D2)). These antibodies specifically immunoprecipitated and recognized by Western blotting RAR-γ1 proteins in COS-1 cells transfected with expression vectors containing the RAR-γ1 cDNAs. They all reacted with both human and mouse RAR-γ1 proteins, except Ab4γ(hF) that was specific for hRAR-γ1. Rabbit polyclonal antibodies, directed against a peptide from the mRAR-γ1 F region were also obtained (RPγ(mF)) and found to be specific for mouse RAR-γ1 protein. Furthermore, in gel retardation/shift assays the antibodies specifically retarded the migration of complexes obtained with a RA response element (RARE). Antibodies raised against regions D2 and F also recognized the RAR-γ2 isoform which differs from RAR-γ1 only in the A region. On the other hand, antibodies directed against the A1 region of RAR-γ1 (Ab1γ(A1)) only reacted with the RAR-γ1 protein. The antibodies characterized here allowed us to detect the presence of mRAR-γ1 and γ2 isoforms in mouse embryos and F9 embryonal carcinoma cells nuclear extracts. They were also used to demonstrate that the mRAR-γ1 protein can be phosphorylated and that the phosphorylation occurs mainly in the NH2-terminal A/B region. |
| doi_str_mv | 10.1083/jcb.115.2.535 |
| format | Article |
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Gaub ; Chambon, P.</creator><creatorcontrib>Rochette-Egly, C. ; Lutz, Y. ; Saunders, M. ; Scheuer, I. ; M.-P. Gaub ; Chambon, P.</creatorcontrib><description>Synthetic peptides corresponding to cDNA-deduced amino acid sequences unique to the human and mouse retinoic acid receptor γ1 (hRAR-γ1 and mRAR-γ1, respectively) were used to generate anti-RAR-γ1 antibodies. Four mAbs were selected, which were directed against peptides found in region A1 (Ab1γ(A1)), region F (Ab2γ(mF) and Ab4γ(hF)) and region D2 (Ab5γ(D2)). These antibodies specifically immunoprecipitated and recognized by Western blotting RAR-γ1 proteins in COS-1 cells transfected with expression vectors containing the RAR-γ1 cDNAs. They all reacted with both human and mouse RAR-γ1 proteins, except Ab4γ(hF) that was specific for hRAR-γ1. Rabbit polyclonal antibodies, directed against a peptide from the mRAR-γ1 F region were also obtained (RPγ(mF)) and found to be specific for mouse RAR-γ1 protein. Furthermore, in gel retardation/shift assays the antibodies specifically retarded the migration of complexes obtained with a RA response element (RARE). Antibodies raised against regions D2 and F also recognized the RAR-γ2 isoform which differs from RAR-γ1 only in the A region. On the other hand, antibodies directed against the A1 region of RAR-γ1 (Ab1γ(A1)) only reacted with the RAR-γ1 protein. The antibodies characterized here allowed us to detect the presence of mRAR-γ1 and γ2 isoforms in mouse embryos and F9 embryonal carcinoma cells nuclear extracts. They were also used to demonstrate that the mRAR-γ1 protein can be phosphorylated and that the phosphorylation occurs mainly in the NH2-terminal A/B region.</description><identifier>ISSN: 0021-9525</identifier><identifier>EISSN: 1540-8140</identifier><identifier>DOI: 10.1083/jcb.115.2.535</identifier><identifier>PMID: 1655807</identifier><identifier>CODEN: JCLBA3</identifier><language>eng</language><publisher>New York, NY: Rockefeller University Press</publisher><subject>Alkaline Phosphatase - metabolism ; Amino Acid Sequence ; Amino acids ; Animals ; Antibodies ; Antibodies, Monoclonal ; Biological and medical sciences ; Blotting, Western ; Carrier Proteins - analysis ; Carrier Proteins - immunology ; Carrier Proteins - metabolism ; Cell extracts ; Cloning, Molecular ; COS cells ; Embryo, Mammalian - metabolism ; Embryos ; F layer ; Fundamental and applied biological sciences. Psychology ; Gels ; Humans ; Mice ; Molecular and cellular biology ; Molecular genetics ; Molecular Sequence Data ; Peptide Fragments - immunology ; Phosphorylation ; Protein isoforms ; Protein Processing, Post-Translational - physiology ; Rabbits ; Receptors, Retinoic Acid ; Retinoic acid receptors ; Transfection - genetics ; Tretinoin - metabolism ; Tumor Cells, Cultured</subject><ispartof>The Journal of cell biology, 1991-10, Vol.115 (2), p.535-545</ispartof><rights>Copyright 1991 The Rockefeller University Press</rights><rights>1992 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c432t-14b17e4d46bb4b64a6938e27ae303a954f633e1560bfdb60d095c37ca852bce53</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,780,784,885,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=5212297$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1655807$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Rochette-Egly, C.</creatorcontrib><creatorcontrib>Lutz, Y.</creatorcontrib><creatorcontrib>Saunders, M.</creatorcontrib><creatorcontrib>Scheuer, I.</creatorcontrib><creatorcontrib>M.-P. Gaub</creatorcontrib><creatorcontrib>Chambon, P.</creatorcontrib><title>Retinoic Acid Receptor γ: Specific Immunodetection and Phosphorylation</title><title>The Journal of cell biology</title><addtitle>J Cell Biol</addtitle><description>Synthetic peptides corresponding to cDNA-deduced amino acid sequences unique to the human and mouse retinoic acid receptor γ1 (hRAR-γ1 and mRAR-γ1, respectively) were used to generate anti-RAR-γ1 antibodies. Four mAbs were selected, which were directed against peptides found in region A1 (Ab1γ(A1)), region F (Ab2γ(mF) and Ab4γ(hF)) and region D2 (Ab5γ(D2)). These antibodies specifically immunoprecipitated and recognized by Western blotting RAR-γ1 proteins in COS-1 cells transfected with expression vectors containing the RAR-γ1 cDNAs. They all reacted with both human and mouse RAR-γ1 proteins, except Ab4γ(hF) that was specific for hRAR-γ1. Rabbit polyclonal antibodies, directed against a peptide from the mRAR-γ1 F region were also obtained (RPγ(mF)) and found to be specific for mouse RAR-γ1 protein. Furthermore, in gel retardation/shift assays the antibodies specifically retarded the migration of complexes obtained with a RA response element (RARE). Antibodies raised against regions D2 and F also recognized the RAR-γ2 isoform which differs from RAR-γ1 only in the A region. On the other hand, antibodies directed against the A1 region of RAR-γ1 (Ab1γ(A1)) only reacted with the RAR-γ1 protein. The antibodies characterized here allowed us to detect the presence of mRAR-γ1 and γ2 isoforms in mouse embryos and F9 embryonal carcinoma cells nuclear extracts. They were also used to demonstrate that the mRAR-γ1 protein can be phosphorylated and that the phosphorylation occurs mainly in the NH2-terminal A/B region.</description><subject>Alkaline Phosphatase - metabolism</subject><subject>Amino Acid Sequence</subject><subject>Amino acids</subject><subject>Animals</subject><subject>Antibodies</subject><subject>Antibodies, Monoclonal</subject><subject>Biological and medical sciences</subject><subject>Blotting, Western</subject><subject>Carrier Proteins - analysis</subject><subject>Carrier Proteins - immunology</subject><subject>Carrier Proteins - metabolism</subject><subject>Cell extracts</subject><subject>Cloning, Molecular</subject><subject>COS cells</subject><subject>Embryo, Mammalian - metabolism</subject><subject>Embryos</subject><subject>F layer</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gels</subject><subject>Humans</subject><subject>Mice</subject><subject>Molecular and cellular biology</subject><subject>Molecular genetics</subject><subject>Molecular Sequence Data</subject><subject>Peptide Fragments - immunology</subject><subject>Phosphorylation</subject><subject>Protein isoforms</subject><subject>Protein Processing, Post-Translational - physiology</subject><subject>Rabbits</subject><subject>Receptors, Retinoic Acid</subject><subject>Retinoic acid receptors</subject><subject>Transfection - genetics</subject><subject>Tretinoin - metabolism</subject><subject>Tumor Cells, Cultured</subject><issn>0021-9525</issn><issn>1540-8140</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1991</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkctKxDAUhoMoOo4u3Sl0Ie465to2LgQRbyAoXtYhSU-dDG1Tk47gc_kePpMdZtBxFTj_x3_CdxA6IHhCcMFOZ9ZMCBETOhFMbKARERynBeF4E40wpiSVgoodtBvjDGPMc8620TbJhChwPkI3T9C71jubXFhXJk9goet9SL6_zpLnDqyrhuiuaeatL6EH2zvfJrotk8epj93Uh89aL2Z7aKvSdYT91TtGr9dXL5e36f3Dzd3lxX1qOaN9SrghOfCSZ8Zwk3GdSVYAzTUwzLQUvMoYAyIybKrSZLjEUliWW10IaiwINkbny95ubhooLbR90LXqgmt0-FReO_U_ad1UvfkPRWkhiVgUnKwKgn-fQ-xV46KFutYt-HlUOSVMyiIfwHQJ2uBjDFD9LiFYLcyrwbwazCuqBvMDf7T-sz96qXrIj1e5jlbXVdCtdfEXE5RQKhfY4RKbxeEOay2EZ7JgPyr9lvI</recordid><startdate>19911001</startdate><enddate>19911001</enddate><creator>Rochette-Egly, C.</creator><creator>Lutz, Y.</creator><creator>Saunders, M.</creator><creator>Scheuer, I.</creator><creator>M.-P. Gaub</creator><creator>Chambon, P.</creator><general>Rockefeller University Press</general><general>The Rockefeller University Press</general><scope>IQODW</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>5PM</scope></search><sort><creationdate>19911001</creationdate><title>Retinoic Acid Receptor γ: Specific Immunodetection and Phosphorylation</title><author>Rochette-Egly, C. ; Lutz, Y. ; Saunders, M. ; Scheuer, I. ; M.-P. Gaub ; Chambon, P.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c432t-14b17e4d46bb4b64a6938e27ae303a954f633e1560bfdb60d095c37ca852bce53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1991</creationdate><topic>Alkaline Phosphatase - metabolism</topic><topic>Amino Acid Sequence</topic><topic>Amino acids</topic><topic>Animals</topic><topic>Antibodies</topic><topic>Antibodies, Monoclonal</topic><topic>Biological and medical sciences</topic><topic>Blotting, Western</topic><topic>Carrier Proteins - analysis</topic><topic>Carrier Proteins - immunology</topic><topic>Carrier Proteins - metabolism</topic><topic>Cell extracts</topic><topic>Cloning, Molecular</topic><topic>COS cells</topic><topic>Embryo, Mammalian - metabolism</topic><topic>Embryos</topic><topic>F layer</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gels</topic><topic>Humans</topic><topic>Mice</topic><topic>Molecular and cellular biology</topic><topic>Molecular genetics</topic><topic>Molecular Sequence Data</topic><topic>Peptide Fragments - immunology</topic><topic>Phosphorylation</topic><topic>Protein isoforms</topic><topic>Protein Processing, Post-Translational - physiology</topic><topic>Rabbits</topic><topic>Receptors, Retinoic Acid</topic><topic>Retinoic acid receptors</topic><topic>Transfection - genetics</topic><topic>Tretinoin - metabolism</topic><topic>Tumor Cells, Cultured</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Rochette-Egly, C.</creatorcontrib><creatorcontrib>Lutz, Y.</creatorcontrib><creatorcontrib>Saunders, M.</creatorcontrib><creatorcontrib>Scheuer, I.</creatorcontrib><creatorcontrib>M.-P. Gaub</creatorcontrib><creatorcontrib>Chambon, P.</creatorcontrib><collection>Pascal-Francis</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>PubMed Central (Full Participant titles)</collection><jtitle>The Journal of cell biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Rochette-Egly, C.</au><au>Lutz, Y.</au><au>Saunders, M.</au><au>Scheuer, I.</au><au>M.-P. Gaub</au><au>Chambon, P.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Retinoic Acid Receptor γ: Specific Immunodetection and Phosphorylation</atitle><jtitle>The Journal of cell biology</jtitle><addtitle>J Cell Biol</addtitle><date>1991-10-01</date><risdate>1991</risdate><volume>115</volume><issue>2</issue><spage>535</spage><epage>545</epage><pages>535-545</pages><issn>0021-9525</issn><eissn>1540-8140</eissn><coden>JCLBA3</coden><abstract>Synthetic peptides corresponding to cDNA-deduced amino acid sequences unique to the human and mouse retinoic acid receptor γ1 (hRAR-γ1 and mRAR-γ1, respectively) were used to generate anti-RAR-γ1 antibodies. Four mAbs were selected, which were directed against peptides found in region A1 (Ab1γ(A1)), region F (Ab2γ(mF) and Ab4γ(hF)) and region D2 (Ab5γ(D2)). These antibodies specifically immunoprecipitated and recognized by Western blotting RAR-γ1 proteins in COS-1 cells transfected with expression vectors containing the RAR-γ1 cDNAs. They all reacted with both human and mouse RAR-γ1 proteins, except Ab4γ(hF) that was specific for hRAR-γ1. Rabbit polyclonal antibodies, directed against a peptide from the mRAR-γ1 F region were also obtained (RPγ(mF)) and found to be specific for mouse RAR-γ1 protein. Furthermore, in gel retardation/shift assays the antibodies specifically retarded the migration of complexes obtained with a RA response element (RARE). Antibodies raised against regions D2 and F also recognized the RAR-γ2 isoform which differs from RAR-γ1 only in the A region. On the other hand, antibodies directed against the A1 region of RAR-γ1 (Ab1γ(A1)) only reacted with the RAR-γ1 protein. The antibodies characterized here allowed us to detect the presence of mRAR-γ1 and γ2 isoforms in mouse embryos and F9 embryonal carcinoma cells nuclear extracts. They were also used to demonstrate that the mRAR-γ1 protein can be phosphorylated and that the phosphorylation occurs mainly in the NH2-terminal A/B region.</abstract><cop>New York, NY</cop><pub>Rockefeller University Press</pub><pmid>1655807</pmid><doi>10.1083/jcb.115.2.535</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | The Journal of cell biology, 1991-10, Vol.115 (2), p.535-545 |
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| language | eng |
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| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection |
| subjects | Alkaline Phosphatase - metabolism Amino Acid Sequence Amino acids Animals Antibodies Antibodies, Monoclonal Biological and medical sciences Blotting, Western Carrier Proteins - analysis Carrier Proteins - immunology Carrier Proteins - metabolism Cell extracts Cloning, Molecular COS cells Embryo, Mammalian - metabolism Embryos F layer Fundamental and applied biological sciences. Psychology Gels Humans Mice Molecular and cellular biology Molecular genetics Molecular Sequence Data Peptide Fragments - immunology Phosphorylation Protein isoforms Protein Processing, Post-Translational - physiology Rabbits Receptors, Retinoic Acid Retinoic acid receptors Transfection - genetics Tretinoin - metabolism Tumor Cells, Cultured |
| title | Retinoic Acid Receptor γ: Specific Immunodetection and Phosphorylation |
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