The Hepatitis Delta Virus Large Antigen Is Farnesylated Both in Vitro and in Animal Cells
The hepatitis delta virus large antigen (lHDAg) is a virally encoded protein that contains a prenylation signal sequence at its carboxyl terminus consisting of the tetrapeptide Cys-Arg-Pro-Gln. Although the presence of the Gln as the COOH-terminal residue generally specifies addition of the 15-carbo...
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Veröffentlicht in: | The Journal of biological chemistry 1996-03, Vol.271 (9), p.4569-4572 |
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description | The hepatitis delta virus large antigen (lHDAg) is a virally encoded protein that contains a prenylation signal sequence at
its carboxyl terminus consisting of the tetrapeptide Cys-Arg-Pro-Gln. Although the presence of the Gln as the COOH-terminal
residue generally specifies addition of the 15-carbon farnesyl isoprenoid, earlier reports had suggested that the protein
is modified by the 20-carbon geranylgeranyl. The prenylation of lHDAg was examined in vitro using a fusion protein between glutathione S -transferase and the COOH-terminal 117 amino acids of lHDAg (GST-lHDAg). When recombinant GST-lHDAg was incubated with bovine
brain cytosol in the presence of either farnesyl diphosphate or geranylgeranyl diphosphate, GST-lHDAg was preferentially farnesylated.
Geranylgeranylation of the fusion protein was also observed, although at a rate considerably less than that of farnesylation.
Using purified recombinant protein prenyltransferases, GST-lHDAg was found to be an excellent substrate (apparent K = 0.8 μM) for protein farnesyltransferase (FTase), while modification by protein geranylgeranyltransferase I (GGTase I) was
not detected. FTase was also able to catalyze geranylgeranylation of GST-lHDAg at a very low rate, suggesting that the low
level of geranylgeranylation of GST-lHDAg observed in cytosolic preparations was mediated by FTase. Consistent with our observations
on the in vitro prenylation of the GST-lHDAg fusion protein, isoprenoid analysis of authentic lHDAg expressed in COS cells demonstrated that
the protein was farnesylated. Geranylgeranylation of lHDAg expressed in COS cells was not observed. As prenylation of lHDAg
is required for the assembly of the hepatitis delta viral particle, these results suggest that inhibitors of FTase may be
useful therapeutic agents for treatment of delta virus infection. |
doi_str_mv | 10.1074/jbc.271.9.4569 |
format | Article |
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its carboxyl terminus consisting of the tetrapeptide Cys-Arg-Pro-Gln. Although the presence of the Gln as the COOH-terminal
residue generally specifies addition of the 15-carbon farnesyl isoprenoid, earlier reports had suggested that the protein
is modified by the 20-carbon geranylgeranyl. The prenylation of lHDAg was examined in vitro using a fusion protein between glutathione S -transferase and the COOH-terminal 117 amino acids of lHDAg (GST-lHDAg). When recombinant GST-lHDAg was incubated with bovine
brain cytosol in the presence of either farnesyl diphosphate or geranylgeranyl diphosphate, GST-lHDAg was preferentially farnesylated.
Geranylgeranylation of the fusion protein was also observed, although at a rate considerably less than that of farnesylation.
Using purified recombinant protein prenyltransferases, GST-lHDAg was found to be an excellent substrate (apparent K = 0.8 μM) for protein farnesyltransferase (FTase), while modification by protein geranylgeranyltransferase I (GGTase I) was
not detected. FTase was also able to catalyze geranylgeranylation of GST-lHDAg at a very low rate, suggesting that the low
level of geranylgeranylation of GST-lHDAg observed in cytosolic preparations was mediated by FTase. Consistent with our observations
on the in vitro prenylation of the GST-lHDAg fusion protein, isoprenoid analysis of authentic lHDAg expressed in COS cells demonstrated that
the protein was farnesylated. Geranylgeranylation of lHDAg expressed in COS cells was not observed. As prenylation of lHDAg
is required for the assembly of the hepatitis delta viral particle, these results suggest that inhibitors of FTase may be
useful therapeutic agents for treatment of delta virus infection.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1074/jbc.271.9.4569</identifier><identifier>PMID: 8617711</identifier><language>eng</language><publisher>United States: American Society for Biochemistry and Molecular Biology</publisher><subject>Amino Acid Sequence ; Animals ; Brain - enzymology ; Cattle ; Cell Line ; Chlorocebus aethiops ; Chromatography, High Pressure Liquid ; Cytosol - enzymology ; Dimethylallyltranstransferase - metabolism ; Glutathione Transferase - metabolism ; Hepatitis Antigens - chemistry ; Hepatitis Antigens - isolation & purification ; Hepatitis Antigens - metabolism ; hepatitis D virus ; Hepatitis delta Antigens ; Hepatitis Delta Virus - metabolism ; Kinetics ; Molecular Sequence Data ; Polyisoprenyl Phosphates - metabolism ; Protein Prenylation ; Recombinant Fusion Proteins - isolation & purification ; Recombinant Fusion Proteins - metabolism ; Sesquiterpenes ; Spodoptera ; Transfection</subject><ispartof>The Journal of biological chemistry, 1996-03, Vol.271 (9), p.4569-4572</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c388t-51b29366fa33f4f2e33d7ce1437ec405e563b95aaeffe25e74b1f330774f65053</citedby><cites>FETCH-LOGICAL-c388t-51b29366fa33f4f2e33d7ce1437ec405e563b95aaeffe25e74b1f330774f65053</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8617711$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Otto, J C</creatorcontrib><creatorcontrib>Casey, P J</creatorcontrib><title>The Hepatitis Delta Virus Large Antigen Is Farnesylated Both in Vitro and in Animal Cells</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>The hepatitis delta virus large antigen (lHDAg) is a virally encoded protein that contains a prenylation signal sequence at
its carboxyl terminus consisting of the tetrapeptide Cys-Arg-Pro-Gln. Although the presence of the Gln as the COOH-terminal
residue generally specifies addition of the 15-carbon farnesyl isoprenoid, earlier reports had suggested that the protein
is modified by the 20-carbon geranylgeranyl. The prenylation of lHDAg was examined in vitro using a fusion protein between glutathione S -transferase and the COOH-terminal 117 amino acids of lHDAg (GST-lHDAg). When recombinant GST-lHDAg was incubated with bovine
brain cytosol in the presence of either farnesyl diphosphate or geranylgeranyl diphosphate, GST-lHDAg was preferentially farnesylated.
Geranylgeranylation of the fusion protein was also observed, although at a rate considerably less than that of farnesylation.
Using purified recombinant protein prenyltransferases, GST-lHDAg was found to be an excellent substrate (apparent K = 0.8 μM) for protein farnesyltransferase (FTase), while modification by protein geranylgeranyltransferase I (GGTase I) was
not detected. FTase was also able to catalyze geranylgeranylation of GST-lHDAg at a very low rate, suggesting that the low
level of geranylgeranylation of GST-lHDAg observed in cytosolic preparations was mediated by FTase. Consistent with our observations
on the in vitro prenylation of the GST-lHDAg fusion protein, isoprenoid analysis of authentic lHDAg expressed in COS cells demonstrated that
the protein was farnesylated. Geranylgeranylation of lHDAg expressed in COS cells was not observed. As prenylation of lHDAg
is required for the assembly of the hepatitis delta viral particle, these results suggest that inhibitors of FTase may be
useful therapeutic agents for treatment of delta virus infection.</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Brain - enzymology</subject><subject>Cattle</subject><subject>Cell Line</subject><subject>Chlorocebus aethiops</subject><subject>Chromatography, High Pressure Liquid</subject><subject>Cytosol - enzymology</subject><subject>Dimethylallyltranstransferase - metabolism</subject><subject>Glutathione Transferase - metabolism</subject><subject>Hepatitis Antigens - chemistry</subject><subject>Hepatitis Antigens - isolation & purification</subject><subject>Hepatitis Antigens - metabolism</subject><subject>hepatitis D virus</subject><subject>Hepatitis delta Antigens</subject><subject>Hepatitis Delta Virus - metabolism</subject><subject>Kinetics</subject><subject>Molecular Sequence Data</subject><subject>Polyisoprenyl Phosphates - metabolism</subject><subject>Protein Prenylation</subject><subject>Recombinant Fusion Proteins - isolation & purification</subject><subject>Recombinant Fusion Proteins - metabolism</subject><subject>Sesquiterpenes</subject><subject>Spodoptera</subject><subject>Transfection</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkMFr2zAUh8Xo6LJu190Goofd7OpJlmUfs3RpCoFd2rKdhOw8xQqOnUkypf_9FBJKb30X8fh9-iF9hHwDlgNTxc2uaXOuIK_zQpb1BzIDVolMSPhzQWaMcchqLqtP5HMIO5amqOGSXFYlKAUwI38fOqQrPJjoogv0Fvto6JPzU6Br47dI50N0WxzofaBL4wcML72JuKE_x9hRNyQ2-pGaYXNc5oPbm54usO_DF_LRmj7g1_N5RR6Xvx4Wq2z9--5-MV9nraiqmEloeC3K0hohbGE5CrFRLUIhFLYFkyhL0dTSGLQWuURVNGCFYEoVtpRMiivy49R78OO_CUPUexfa9AIz4DgFrSrGFQf-LgiKlZUsIYH5CWz9GIJHqw8-_cu_aGD6KF0n6TpJ17U-Sk8Xvp-bp2aPm1f8bDnl16e8c9vu2XnUjRvbDvdvS_4D2z-HNg</recordid><startdate>19960301</startdate><enddate>19960301</enddate><creator>Otto, J C</creator><creator>Casey, P J</creator><general>American Society for Biochemistry and Molecular Biology</general><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>7U9</scope><scope>H94</scope><scope>7X8</scope></search><sort><creationdate>19960301</creationdate><title>The Hepatitis Delta Virus Large Antigen Is Farnesylated Both in Vitro and in Animal Cells</title><author>Otto, J C ; Casey, P J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c388t-51b29366fa33f4f2e33d7ce1437ec405e563b95aaeffe25e74b1f330774f65053</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1996</creationdate><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Brain - enzymology</topic><topic>Cattle</topic><topic>Cell Line</topic><topic>Chlorocebus aethiops</topic><topic>Chromatography, High Pressure Liquid</topic><topic>Cytosol - enzymology</topic><topic>Dimethylallyltranstransferase - metabolism</topic><topic>Glutathione Transferase - metabolism</topic><topic>Hepatitis Antigens - chemistry</topic><topic>Hepatitis Antigens - isolation & purification</topic><topic>Hepatitis Antigens - metabolism</topic><topic>hepatitis D virus</topic><topic>Hepatitis delta Antigens</topic><topic>Hepatitis Delta Virus - metabolism</topic><topic>Kinetics</topic><topic>Molecular Sequence Data</topic><topic>Polyisoprenyl Phosphates - metabolism</topic><topic>Protein Prenylation</topic><topic>Recombinant Fusion Proteins - isolation & purification</topic><topic>Recombinant Fusion Proteins - metabolism</topic><topic>Sesquiterpenes</topic><topic>Spodoptera</topic><topic>Transfection</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Otto, J C</creatorcontrib><creatorcontrib>Casey, P J</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Otto, J C</au><au>Casey, P J</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The Hepatitis Delta Virus Large Antigen Is Farnesylated Both in Vitro and in Animal Cells</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>1996-03-01</date><risdate>1996</risdate><volume>271</volume><issue>9</issue><spage>4569</spage><epage>4572</epage><pages>4569-4572</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>The hepatitis delta virus large antigen (lHDAg) is a virally encoded protein that contains a prenylation signal sequence at
its carboxyl terminus consisting of the tetrapeptide Cys-Arg-Pro-Gln. Although the presence of the Gln as the COOH-terminal
residue generally specifies addition of the 15-carbon farnesyl isoprenoid, earlier reports had suggested that the protein
is modified by the 20-carbon geranylgeranyl. The prenylation of lHDAg was examined in vitro using a fusion protein between glutathione S -transferase and the COOH-terminal 117 amino acids of lHDAg (GST-lHDAg). When recombinant GST-lHDAg was incubated with bovine
brain cytosol in the presence of either farnesyl diphosphate or geranylgeranyl diphosphate, GST-lHDAg was preferentially farnesylated.
Geranylgeranylation of the fusion protein was also observed, although at a rate considerably less than that of farnesylation.
Using purified recombinant protein prenyltransferases, GST-lHDAg was found to be an excellent substrate (apparent K = 0.8 μM) for protein farnesyltransferase (FTase), while modification by protein geranylgeranyltransferase I (GGTase I) was
not detected. FTase was also able to catalyze geranylgeranylation of GST-lHDAg at a very low rate, suggesting that the low
level of geranylgeranylation of GST-lHDAg observed in cytosolic preparations was mediated by FTase. Consistent with our observations
on the in vitro prenylation of the GST-lHDAg fusion protein, isoprenoid analysis of authentic lHDAg expressed in COS cells demonstrated that
the protein was farnesylated. Geranylgeranylation of lHDAg expressed in COS cells was not observed. As prenylation of lHDAg
is required for the assembly of the hepatitis delta viral particle, these results suggest that inhibitors of FTase may be
useful therapeutic agents for treatment of delta virus infection.</abstract><cop>United States</cop><pub>American Society for Biochemistry and Molecular Biology</pub><pmid>8617711</pmid><doi>10.1074/jbc.271.9.4569</doi><tpages>4</tpages><oa>free_for_read</oa></addata></record> |
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source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection |
subjects | Amino Acid Sequence Animals Brain - enzymology Cattle Cell Line Chlorocebus aethiops Chromatography, High Pressure Liquid Cytosol - enzymology Dimethylallyltranstransferase - metabolism Glutathione Transferase - metabolism Hepatitis Antigens - chemistry Hepatitis Antigens - isolation & purification Hepatitis Antigens - metabolism hepatitis D virus Hepatitis delta Antigens Hepatitis Delta Virus - metabolism Kinetics Molecular Sequence Data Polyisoprenyl Phosphates - metabolism Protein Prenylation Recombinant Fusion Proteins - isolation & purification Recombinant Fusion Proteins - metabolism Sesquiterpenes Spodoptera Transfection |
title | The Hepatitis Delta Virus Large Antigen Is Farnesylated Both in Vitro and in Animal Cells |
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