Expression of human DNA topoisomerase I in yeast cells lacking yeast DNA topoisomerase I: restoration of sensitivity of the cells to the antitumor drug camptothecin
Yeast Saccharomyces cerevisiae strains that are permeable to the antitumor alkaloid camptothecin are killed by the drug if they express DNA topoisomerase I, the cellular target of the drug (J. Nitiss and J.C. Wang, Proc. Natl. Acad. Sci. USA, 85: 7501-7505, 1988). We show that in a yeast strain perm...
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| Veröffentlicht in: | Cancer research (Chicago, Ill.) Ill.), 1989-11, Vol.49 (22), p.6318-6323 |
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| creator | BJORNSTI, M.-A BENEDETTI, P VIGLIANTI, G. A WANG, J. C |
| description | Yeast Saccharomyces cerevisiae strains that are permeable to the antitumor alkaloid camptothecin are killed by the drug if they express DNA topoisomerase I, the cellular target of the drug (J. Nitiss and J.C. Wang, Proc. Natl. Acad. Sci. USA, 85: 7501-7505, 1988). We show that in a yeast strain permeable to camptothecin but lacking DNA topoisomerase I, sensitivity to the drug was restored upon expression of human DNA topoisomerase I from a plasmid-borne human complementary DNA clone. When the human enzyme was expressed from a galactose-inducible, glucose-repressible yeast promoter, PGAL1, sensitivity to camptothecin was observed in the presence of galactose but not in the presence of glucose. Expression of human DNA topoisomerase I in Escherichia coli was also demonstrated by the complementation of a conditional lethal E. coli DNA topoisomerase I mutant. These systems can be used in the study of human DNA topoisomerase I-camptothecin interactions and in the screening of additional therapeutics targeting the enzyme. |
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A ; WANG, J. C</creator><creatorcontrib>BJORNSTI, M.-A ; BENEDETTI, P ; VIGLIANTI, G. A ; WANG, J. C</creatorcontrib><description>Yeast Saccharomyces cerevisiae strains that are permeable to the antitumor alkaloid camptothecin are killed by the drug if they express DNA topoisomerase I, the cellular target of the drug (J. Nitiss and J.C. Wang, Proc. Natl. Acad. Sci. USA, 85: 7501-7505, 1988). We show that in a yeast strain permeable to camptothecin but lacking DNA topoisomerase I, sensitivity to the drug was restored upon expression of human DNA topoisomerase I from a plasmid-borne human complementary DNA clone. When the human enzyme was expressed from a galactose-inducible, glucose-repressible yeast promoter, PGAL1, sensitivity to camptothecin was observed in the presence of galactose but not in the presence of glucose. Expression of human DNA topoisomerase I in Escherichia coli was also demonstrated by the complementation of a conditional lethal E. coli DNA topoisomerase I mutant. These systems can be used in the study of human DNA topoisomerase I-camptothecin interactions and in the screening of additional therapeutics targeting the enzyme.</description><identifier>ISSN: 0008-5472</identifier><identifier>EISSN: 1538-7445</identifier><identifier>PMID: 2553253</identifier><identifier>CODEN: CNREA8</identifier><language>eng</language><publisher>Philadelphia, PA: American Association for Cancer Research</publisher><subject>Antineoplastic agents ; Base Sequence ; Biological and medical sciences ; Camptothecin - metabolism ; Camptothecin - pharmacology ; DNA Topoisomerases, Type I - genetics ; DNA Topoisomerases, Type I - metabolism ; Escherichia coli - enzymology ; Escherichia coli - genetics ; General aspects ; Genes ; Genetic Complementation Test ; HeLa Cells - enzymology ; Humans ; Kinetics ; Medical sciences ; Molecular Sequence Data ; Pharmacology. Drug treatments ; Plasmids ; Recombinant Proteins - metabolism ; Saccharomyces cerevisiae ; Saccharomyces cerevisiae - drug effects ; Saccharomyces cerevisiae - enzymology ; Saccharomyces cerevisiae - genetics</subject><ispartof>Cancer research (Chicago, Ill.), 1989-11, Vol.49 (22), p.6318-6323</ispartof><rights>1990 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=6655391$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2553253$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>BJORNSTI, M.-A</creatorcontrib><creatorcontrib>BENEDETTI, P</creatorcontrib><creatorcontrib>VIGLIANTI, G. A</creatorcontrib><creatorcontrib>WANG, J. C</creatorcontrib><title>Expression of human DNA topoisomerase I in yeast cells lacking yeast DNA topoisomerase I: restoration of sensitivity of the cells to the antitumor drug camptothecin</title><title>Cancer research (Chicago, Ill.)</title><addtitle>Cancer Res</addtitle><description>Yeast Saccharomyces cerevisiae strains that are permeable to the antitumor alkaloid camptothecin are killed by the drug if they express DNA topoisomerase I, the cellular target of the drug (J. Nitiss and J.C. Wang, Proc. Natl. Acad. Sci. USA, 85: 7501-7505, 1988). We show that in a yeast strain permeable to camptothecin but lacking DNA topoisomerase I, sensitivity to the drug was restored upon expression of human DNA topoisomerase I from a plasmid-borne human complementary DNA clone. When the human enzyme was expressed from a galactose-inducible, glucose-repressible yeast promoter, PGAL1, sensitivity to camptothecin was observed in the presence of galactose but not in the presence of glucose. Expression of human DNA topoisomerase I in Escherichia coli was also demonstrated by the complementation of a conditional lethal E. coli DNA topoisomerase I mutant. These systems can be used in the study of human DNA topoisomerase I-camptothecin interactions and in the screening of additional therapeutics targeting the enzyme.</description><subject>Antineoplastic agents</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Camptothecin - metabolism</subject><subject>Camptothecin - pharmacology</subject><subject>DNA Topoisomerases, Type I - genetics</subject><subject>DNA Topoisomerases, Type I - metabolism</subject><subject>Escherichia coli - enzymology</subject><subject>Escherichia coli - genetics</subject><subject>General aspects</subject><subject>Genes</subject><subject>Genetic Complementation Test</subject><subject>HeLa Cells - enzymology</subject><subject>Humans</subject><subject>Kinetics</subject><subject>Medical sciences</subject><subject>Molecular Sequence Data</subject><subject>Pharmacology. Drug treatments</subject><subject>Plasmids</subject><subject>Recombinant Proteins - metabolism</subject><subject>Saccharomyces cerevisiae</subject><subject>Saccharomyces cerevisiae - drug effects</subject><subject>Saccharomyces cerevisiae - enzymology</subject><subject>Saccharomyces cerevisiae - genetics</subject><issn>0008-5472</issn><issn>1538-7445</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1989</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkFFPwyAUhRujmXP6E0x4ML41gQJt8W2ZU5cYfdHn5q6FDW2hAjXu__hDZdr4ZOJ9gcP5ODm5B8mUcFqmBWP8MJlijMuUsyI7Tk68f4mSE8wnySTjnGacTpPP5UfvpPfaGmQV2g4dGHT9MEfB9lZ720kHXqIV0gbtJPiAatm2HrVQv2qzGd_--HCFYmywDsIY7aXxOuh3HXZ7GbZyjAr2W4AJOgyddahxwwbV0PXBRqPW5jQ5UtB6eTaes-T5Zvm0uEvvH29Xi_l9uqUYh5SImnJRl6CwWhOWqUwBFbwRjCpWMiokLwjHUKh1HELzeIWcyHXTlIyUmM6Sy5_c3tm3IdavOu33JcFIO_iqEFmBSyr-BQlnGcV5HsHzERzWnWyq3ukO3K4a9x_9i9EHX0OrHJha-18szyMnCP0CVeqR8g</recordid><startdate>19891115</startdate><enddate>19891115</enddate><creator>BJORNSTI, M.-A</creator><creator>BENEDETTI, P</creator><creator>VIGLIANTI, G. A</creator><creator>WANG, J. C</creator><general>American Association for Cancer Research</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7TM</scope><scope>M7N</scope><scope>7X8</scope></search><sort><creationdate>19891115</creationdate><title>Expression of human DNA topoisomerase I in yeast cells lacking yeast DNA topoisomerase I: restoration of sensitivity of the cells to the antitumor drug camptothecin</title><author>BJORNSTI, M.-A ; BENEDETTI, P ; VIGLIANTI, G. A ; WANG, J. C</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-h300t-19c359c8af0fb142f2fa395d943f48439e57150a7fbbbb1360a7a61ebdd841803</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1989</creationdate><topic>Antineoplastic agents</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Camptothecin - metabolism</topic><topic>Camptothecin - pharmacology</topic><topic>DNA Topoisomerases, Type I - genetics</topic><topic>DNA Topoisomerases, Type I - metabolism</topic><topic>Escherichia coli - enzymology</topic><topic>Escherichia coli - genetics</topic><topic>General aspects</topic><topic>Genes</topic><topic>Genetic Complementation Test</topic><topic>HeLa Cells - enzymology</topic><topic>Humans</topic><topic>Kinetics</topic><topic>Medical sciences</topic><topic>Molecular Sequence Data</topic><topic>Pharmacology. Drug treatments</topic><topic>Plasmids</topic><topic>Recombinant Proteins - metabolism</topic><topic>Saccharomyces cerevisiae</topic><topic>Saccharomyces cerevisiae - drug effects</topic><topic>Saccharomyces cerevisiae - enzymology</topic><topic>Saccharomyces cerevisiae - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>BJORNSTI, M.-A</creatorcontrib><creatorcontrib>BENEDETTI, P</creatorcontrib><creatorcontrib>VIGLIANTI, G. A</creatorcontrib><creatorcontrib>WANG, J. C</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>Nucleic Acids Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>MEDLINE - Academic</collection><jtitle>Cancer research (Chicago, Ill.)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>BJORNSTI, M.-A</au><au>BENEDETTI, P</au><au>VIGLIANTI, G. A</au><au>WANG, J. C</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Expression of human DNA topoisomerase I in yeast cells lacking yeast DNA topoisomerase I: restoration of sensitivity of the cells to the antitumor drug camptothecin</atitle><jtitle>Cancer research (Chicago, Ill.)</jtitle><addtitle>Cancer Res</addtitle><date>1989-11-15</date><risdate>1989</risdate><volume>49</volume><issue>22</issue><spage>6318</spage><epage>6323</epage><pages>6318-6323</pages><issn>0008-5472</issn><eissn>1538-7445</eissn><coden>CNREA8</coden><abstract>Yeast Saccharomyces cerevisiae strains that are permeable to the antitumor alkaloid camptothecin are killed by the drug if they express DNA topoisomerase I, the cellular target of the drug (J. Nitiss and J.C. Wang, Proc. Natl. Acad. Sci. USA, 85: 7501-7505, 1988). We show that in a yeast strain permeable to camptothecin but lacking DNA topoisomerase I, sensitivity to the drug was restored upon expression of human DNA topoisomerase I from a plasmid-borne human complementary DNA clone. When the human enzyme was expressed from a galactose-inducible, glucose-repressible yeast promoter, PGAL1, sensitivity to camptothecin was observed in the presence of galactose but not in the presence of glucose. Expression of human DNA topoisomerase I in Escherichia coli was also demonstrated by the complementation of a conditional lethal E. coli DNA topoisomerase I mutant. These systems can be used in the study of human DNA topoisomerase I-camptothecin interactions and in the screening of additional therapeutics targeting the enzyme.</abstract><cop>Philadelphia, PA</cop><pub>American Association for Cancer Research</pub><pmid>2553253</pmid><tpages>6</tpages></addata></record> |
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| source | MEDLINE; American Association for Cancer Research; EZB-FREE-00999 freely available EZB journals |
| subjects | Antineoplastic agents Base Sequence Biological and medical sciences Camptothecin - metabolism Camptothecin - pharmacology DNA Topoisomerases, Type I - genetics DNA Topoisomerases, Type I - metabolism Escherichia coli - enzymology Escherichia coli - genetics General aspects Genes Genetic Complementation Test HeLa Cells - enzymology Humans Kinetics Medical sciences Molecular Sequence Data Pharmacology. Drug treatments Plasmids Recombinant Proteins - metabolism Saccharomyces cerevisiae Saccharomyces cerevisiae - drug effects Saccharomyces cerevisiae - enzymology Saccharomyces cerevisiae - genetics |
| title | Expression of human DNA topoisomerase I in yeast cells lacking yeast DNA topoisomerase I: restoration of sensitivity of the cells to the antitumor drug camptothecin |
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