The 5′ untranslated region of the human cellular glutathione peroxidase gene is indispensable for its expression in COS-7 cells
We studied the expression of the human cellular glutathione peroxidase (GPx) gene, from which a key enzyme containing selenocysteine (Scy) at the active site is produced. Expression of some human GPx gene mutants in COS-7 cells revealed that the 5′ untranslated region (utr) was necessary for express...
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| Veröffentlicht in: | FEBS letters 1992-11, Vol.312 (1), p.10-14 |
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| creator | Kurata, Hiroshi Kamoshita, Keiichi Kawai, Eiko Sukenaga, Yoshikazu Mizutani, Takaharu |
| description | We studied the expression of the human cellular glutathione peroxidase (GPx) gene, from which a key enzyme containing selenocysteine (Scy) at the active site is produced. Expression of some human GPx gene mutants in COS-7 cells revealed that the 5′ untranslated region (utr) was necessary for expression of the GPx gene, since mutant genes having 10 base pairs (bps) at the 5′utr (the complete had 311 bps) expressed GPx at very low levels. The genes with 311 or 408 bps at the 5′utr were better expressed than those having 257 bps. The GPx gene having 133 bps at the 3′utr (80 bps shorter than the entire length) was highly expressed. This deletion did not influence expression. We constructed some mutants in which 3 bases were altered at the upstream region of the Scy UGA codon in the frame of the GPx gene, by site-directed mutagenesis. GPx expression decreased but the expression was restored. Therefore, the upstream region of the in-frame Scy codon was not essential in the Scy decoding mechanisms. Finally, the 5′utr was essential for the expression of GPx gene. However, the deletion of a part of the 3′utr and the site-directed mutation upstream of the Scy codon did not show drastic effects on the expression. |
| doi_str_mv | 10.1016/0014-5793(92)81400-G |
| format | Article |
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Expression of some human GPx gene mutants in COS-7 cells revealed that the 5′ untranslated region (utr) was necessary for expression of the GPx gene, since mutant genes having 10 base pairs (bps) at the 5′utr (the complete had 311 bps) expressed GPx at very low levels. The genes with 311 or 408 bps at the 5′utr were better expressed than those having 257 bps. The GPx gene having 133 bps at the 3′utr (80 bps shorter than the entire length) was highly expressed. This deletion did not influence expression. We constructed some mutants in which 3 bases were altered at the upstream region of the Scy UGA codon in the frame of the GPx gene, by site-directed mutagenesis. GPx expression decreased but the expression was restored. Therefore, the upstream region of the in-frame Scy codon was not essential in the Scy decoding mechanisms. Finally, the 5′utr was essential for the expression of GPx gene. However, the deletion of a part of the 3′utr and the site-directed mutation upstream of the Scy codon did not show drastic effects on the expression.</description><identifier>ISSN: 0014-5793</identifier><identifier>EISSN: 1873-3468</identifier><identifier>DOI: 10.1016/0014-5793(92)81400-G</identifier><identifier>PMID: 1426231</identifier><identifier>CODEN: FEBLAL</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject>5′-iodothyronine monodeiodinase ; 5′-Untranslated region ; 5′DI ; Amino Acid Sequence ; Animals ; Base Composition ; base pairs ; Base Sequence ; Biological and medical sciences ; Blotting, Western ; bps ; Cell Line ; cellular glutathione peroxidase ; dexamethasone ; dxm ; Exons ; FDH ; formate dehydrogenase ; Fundamental and applied biological sciences. Psychology ; Gene Expression ; Genes ; Genetic Vectors ; Glutathione peroxidase ; Glutathione Peroxidase - genetics ; GPx ; Humans ; Introns ; Molecular and cellular biology ; Molecular genetics ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Nucleic Acid Conformation ; PHGPx ; phospholipid hydroperoxide glutathione peroxidase ; plasma selenoprotein ; Protein Biosynthesis ; RNA, Messenger - genetics ; RNA, Messenger - metabolism ; Scy ; Selenium ; Selenocysteine ; SeP ; Site-directed mutagenesis ; Transfection ; untranslated region ; utr</subject><ispartof>FEBS letters, 1992-11, Vol.312 (1), p.10-14</ispartof><rights>1992</rights><rights>FEBS Letters 312 (1992) 1873-3468 © 2015 Federation of European Biochemical Societies</rights><rights>1993 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c480G-101a3c43569cea260f641b81624e60423409014599d5b521490a51086676c13e3</citedby><cites>FETCH-LOGICAL-c480G-101a3c43569cea260f641b81624e60423409014599d5b521490a51086676c13e3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0014-5793(92)81400-G$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>314,777,781,3537,27905,27906,45976</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=4334387$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1426231$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kurata, Hiroshi</creatorcontrib><creatorcontrib>Kamoshita, Keiichi</creatorcontrib><creatorcontrib>Kawai, Eiko</creatorcontrib><creatorcontrib>Sukenaga, Yoshikazu</creatorcontrib><creatorcontrib>Mizutani, Takaharu</creatorcontrib><title>The 5′ untranslated region of the human cellular glutathione peroxidase gene is indispensable for its expression in COS-7 cells</title><title>FEBS letters</title><addtitle>FEBS Lett</addtitle><description>We studied the expression of the human cellular glutathione peroxidase (GPx) gene, from which a key enzyme containing selenocysteine (Scy) at the active site is produced. Expression of some human GPx gene mutants in COS-7 cells revealed that the 5′ untranslated region (utr) was necessary for expression of the GPx gene, since mutant genes having 10 base pairs (bps) at the 5′utr (the complete had 311 bps) expressed GPx at very low levels. The genes with 311 or 408 bps at the 5′utr were better expressed than those having 257 bps. The GPx gene having 133 bps at the 3′utr (80 bps shorter than the entire length) was highly expressed. This deletion did not influence expression. We constructed some mutants in which 3 bases were altered at the upstream region of the Scy UGA codon in the frame of the GPx gene, by site-directed mutagenesis. GPx expression decreased but the expression was restored. Therefore, the upstream region of the in-frame Scy codon was not essential in the Scy decoding mechanisms. Finally, the 5′utr was essential for the expression of GPx gene. However, the deletion of a part of the 3′utr and the site-directed mutation upstream of the Scy codon did not show drastic effects on the expression.</description><subject>5′-iodothyronine monodeiodinase</subject><subject>5′-Untranslated region</subject><subject>5′DI</subject><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Base Composition</subject><subject>base pairs</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Blotting, Western</subject><subject>bps</subject><subject>Cell Line</subject><subject>cellular glutathione peroxidase</subject><subject>dexamethasone</subject><subject>dxm</subject><subject>Exons</subject><subject>FDH</subject><subject>formate dehydrogenase</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression</subject><subject>Genes</subject><subject>Genetic Vectors</subject><subject>Glutathione peroxidase</subject><subject>Glutathione Peroxidase - genetics</subject><subject>GPx</subject><subject>Humans</subject><subject>Introns</subject><subject>Molecular and cellular biology</subject><subject>Molecular genetics</subject><subject>Molecular Sequence Data</subject><subject>Mutagenesis, Site-Directed</subject><subject>Nucleic Acid Conformation</subject><subject>PHGPx</subject><subject>phospholipid hydroperoxide glutathione peroxidase</subject><subject>plasma selenoprotein</subject><subject>Protein Biosynthesis</subject><subject>RNA, Messenger - genetics</subject><subject>RNA, Messenger - metabolism</subject><subject>Scy</subject><subject>Selenium</subject><subject>Selenocysteine</subject><subject>SeP</subject><subject>Site-directed mutagenesis</subject><subject>Transfection</subject><subject>untranslated region</subject><subject>utr</subject><issn>0014-5793</issn><issn>1873-3468</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1992</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkc9u1DAQxi0EKsvCG4DkA0LtIeB_8cYXJFh1l0qVeqCcLa8z2TXKJsGTQHsrr8Qj8SR1NqtyqzhZM9_nzzM_E_Kas_eccf2BMa6yfGHkqRFnBVeMZesnZMaLhcyk0sVTMnuwPCcvEL-zVBfcnJATroQWks_I7-sd0Pzv3R86NH10Ddauh5JG2Ia2oW1F-6Tvhr1rqIe6HmoX6bYeetfvkgFoB7G9CaVDoFtIdUAamjJgBw26TQ20aiMNPVK46SIgjqmhocurr9nikIgvybPK1QivjuecfFudXy-_ZJdX64vlp8vMq4Kts7Syk17JXBsPTmhWacU3BddCgWZKSMVM2jY3psw3ueDKMJdzVmi90J5LkHPybsrtYvtjAOztPuA4gWugHdByLQupeJGMajL62CJGqGwXw97FW8uZHcnbEasdsVoj7IG8Xadrb475w2YP5b9LE-qkvz3qDr2rqwTbB3ywKSmVTF83J6vJ9ivUcPtfT9vV-WcxCmPfiEN3nOfjFAQJ6s8A0aIP0HgoQwTf27INjy90D7Qbsnw</recordid><startdate>19921102</startdate><enddate>19921102</enddate><creator>Kurata, Hiroshi</creator><creator>Kamoshita, Keiichi</creator><creator>Kawai, Eiko</creator><creator>Sukenaga, Yoshikazu</creator><creator>Mizutani, Takaharu</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>6I.</scope><scope>AAFTH</scope><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>7TM</scope></search><sort><creationdate>19921102</creationdate><title>The 5′ untranslated region of the human cellular glutathione peroxidase gene is indispensable for its expression in COS-7 cells</title><author>Kurata, Hiroshi ; Kamoshita, Keiichi ; Kawai, Eiko ; Sukenaga, Yoshikazu ; Mizutani, Takaharu</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c480G-101a3c43569cea260f641b81624e60423409014599d5b521490a51086676c13e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1992</creationdate><topic>5′-iodothyronine monodeiodinase</topic><topic>5′-Untranslated region</topic><topic>5′DI</topic><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Base Composition</topic><topic>base pairs</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Blotting, Western</topic><topic>bps</topic><topic>Cell Line</topic><topic>cellular glutathione peroxidase</topic><topic>dexamethasone</topic><topic>dxm</topic><topic>Exons</topic><topic>FDH</topic><topic>formate dehydrogenase</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression</topic><topic>Genes</topic><topic>Genetic Vectors</topic><topic>Glutathione peroxidase</topic><topic>Glutathione Peroxidase - genetics</topic><topic>GPx</topic><topic>Humans</topic><topic>Introns</topic><topic>Molecular and cellular biology</topic><topic>Molecular genetics</topic><topic>Molecular Sequence Data</topic><topic>Mutagenesis, Site-Directed</topic><topic>Nucleic Acid Conformation</topic><topic>PHGPx</topic><topic>phospholipid hydroperoxide glutathione peroxidase</topic><topic>plasma selenoprotein</topic><topic>Protein Biosynthesis</topic><topic>RNA, Messenger - genetics</topic><topic>RNA, Messenger - metabolism</topic><topic>Scy</topic><topic>Selenium</topic><topic>Selenocysteine</topic><topic>SeP</topic><topic>Site-directed mutagenesis</topic><topic>Transfection</topic><topic>untranslated region</topic><topic>utr</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kurata, Hiroshi</creatorcontrib><creatorcontrib>Kamoshita, Keiichi</creatorcontrib><creatorcontrib>Kawai, Eiko</creatorcontrib><creatorcontrib>Sukenaga, Yoshikazu</creatorcontrib><creatorcontrib>Mizutani, Takaharu</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><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>Nucleic Acids Abstracts</collection><jtitle>FEBS letters</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kurata, Hiroshi</au><au>Kamoshita, Keiichi</au><au>Kawai, Eiko</au><au>Sukenaga, Yoshikazu</au><au>Mizutani, Takaharu</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The 5′ untranslated region of the human cellular glutathione peroxidase gene is indispensable for its expression in COS-7 cells</atitle><jtitle>FEBS letters</jtitle><addtitle>FEBS Lett</addtitle><date>1992-11-02</date><risdate>1992</risdate><volume>312</volume><issue>1</issue><spage>10</spage><epage>14</epage><pages>10-14</pages><issn>0014-5793</issn><eissn>1873-3468</eissn><coden>FEBLAL</coden><abstract>We studied the expression of the human cellular glutathione peroxidase (GPx) gene, from which a key enzyme containing selenocysteine (Scy) at the active site is produced. Expression of some human GPx gene mutants in COS-7 cells revealed that the 5′ untranslated region (utr) was necessary for expression of the GPx gene, since mutant genes having 10 base pairs (bps) at the 5′utr (the complete had 311 bps) expressed GPx at very low levels. The genes with 311 or 408 bps at the 5′utr were better expressed than those having 257 bps. The GPx gene having 133 bps at the 3′utr (80 bps shorter than the entire length) was highly expressed. This deletion did not influence expression. We constructed some mutants in which 3 bases were altered at the upstream region of the Scy UGA codon in the frame of the GPx gene, by site-directed mutagenesis. GPx expression decreased but the expression was restored. Therefore, the upstream region of the in-frame Scy codon was not essential in the Scy decoding mechanisms. Finally, the 5′utr was essential for the expression of GPx gene. However, the deletion of a part of the 3′utr and the site-directed mutation upstream of the Scy codon did not show drastic effects on the expression.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>1426231</pmid><doi>10.1016/0014-5793(92)81400-G</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| identifier | ISSN: 0014-5793 |
| ispartof | FEBS letters, 1992-11, Vol.312 (1), p.10-14 |
| issn | 0014-5793 1873-3468 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_16383418 |
| source | MEDLINE; Elsevier ScienceDirect Journals; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | 5′-iodothyronine monodeiodinase 5′-Untranslated region 5′DI Amino Acid Sequence Animals Base Composition base pairs Base Sequence Biological and medical sciences Blotting, Western bps Cell Line cellular glutathione peroxidase dexamethasone dxm Exons FDH formate dehydrogenase Fundamental and applied biological sciences. Psychology Gene Expression Genes Genetic Vectors Glutathione peroxidase Glutathione Peroxidase - genetics GPx Humans Introns Molecular and cellular biology Molecular genetics Molecular Sequence Data Mutagenesis, Site-Directed Nucleic Acid Conformation PHGPx phospholipid hydroperoxide glutathione peroxidase plasma selenoprotein Protein Biosynthesis RNA, Messenger - genetics RNA, Messenger - metabolism Scy Selenium Selenocysteine SeP Site-directed mutagenesis Transfection untranslated region utr |
| title | The 5′ untranslated region of the human cellular glutathione peroxidase gene is indispensable for its expression in COS-7 cells |
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