M phase-specific association of human topoisomerase IIIbeta with chromosomes
Two isoforms, 1 and 2, of human DNA topoisomerase IIIbeta were expressed in HeLa cells as a fusion protein to the C-terminus of green fluorescent protein (GFP). The fusion protein of the isoform 1 was found to be localized to the nucleus, and to be associated with chromosomes during metaphase and an...
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| Veröffentlicht in: | Biochemical and biophysical research communications 2001-09, Vol.287 (1), p.282-287 |
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| creator | Kobayashi, M Hanai, R |
| description | Two isoforms, 1 and 2, of human DNA topoisomerase IIIbeta were expressed in HeLa cells as a fusion protein to the C-terminus of green fluorescent protein (GFP). The fusion protein of the isoform 1 was found to be localized to the nucleus, and to be associated with chromosomes during metaphase and anaphase. As yeast top3 mutants are known to exhibit phenotypes indicative of defective chromosome segregation, the result suggests that the isoform 1 of the human enzyme may also be involved in chromosome segregation. Two-hybrid screening for interaction partners of the isoform identified three candidate genes: CENP-F, a gene encoding a centromere protein and two genes of no known function, one of which was novel. The GFP fusion of the isoform 2 was found in the cytoplasm, indicating the nuclear localization signal sequence in the isoform 1 is in the C-terminal part that is different between the two isoforms. |
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The fusion protein of the isoform 1 was found to be localized to the nucleus, and to be associated with chromosomes during metaphase and anaphase. As yeast top3 mutants are known to exhibit phenotypes indicative of defective chromosome segregation, the result suggests that the isoform 1 of the human enzyme may also be involved in chromosome segregation. Two-hybrid screening for interaction partners of the isoform identified three candidate genes: CENP-F, a gene encoding a centromere protein and two genes of no known function, one of which was novel. 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The fusion protein of the isoform 1 was found to be localized to the nucleus, and to be associated with chromosomes during metaphase and anaphase. As yeast top3 mutants are known to exhibit phenotypes indicative of defective chromosome segregation, the result suggests that the isoform 1 of the human enzyme may also be involved in chromosome segregation. Two-hybrid screening for interaction partners of the isoform identified three candidate genes: CENP-F, a gene encoding a centromere protein and two genes of no known function, one of which was novel. The GFP fusion of the isoform 2 was found in the cytoplasm, indicating the nuclear localization signal sequence in the isoform 1 is in the C-terminal part that is different between the two isoforms.</description><subject>Amino Acid Sequence</subject><subject>Base Sequence</subject><subject>Chromosomes</subject><subject>DNA Topoisomerases, Type I - genetics</subject><subject>DNA Topoisomerases, Type I - metabolism</subject><subject>Green Fluorescent Proteins</subject><subject>HeLa Cells</subject><subject>Humans</subject><subject>Isoenzymes - genetics</subject><subject>Isoenzymes - metabolism</subject><subject>Luminescent Proteins - metabolism</subject><subject>Mitosis - physiology</subject><subject>Molecular Sequence Data</subject><subject>Subcellular Fractions</subject><issn>0006-291X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2001</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo1j71OwzAYRT2AaCm8AvLEFsl2_j6PqOInUhBLB7bI_mIrRnFs4kSIt6cV7XSHe3Skc0W2jLEqE5J_bshtSl-McV5U8oZsOC8LKQC2pH2ncVDJZCkadNYhVSkFdGpxYaLB0mH1aqJLiMGl4M18ZGnTNNosiv64ZaA4zMGH05fuyLVVYzL3592Rw8vzYf-WtR-vzf6pzWJZQFYA47202mhbqqrOZSXRIuSC8x6FriveAxeATECpJQLYXsocpYUKNDLMd-TxXxvn8L2atHTeJTTjqCYT1tTVpzzI5RF8OIOr9qbv4uy8mn-7S37-BzioVis</recordid><startdate>20010914</startdate><enddate>20010914</enddate><creator>Kobayashi, M</creator><creator>Hanai, R</creator><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope></search><sort><creationdate>20010914</creationdate><title>M phase-specific association of human topoisomerase IIIbeta with chromosomes</title><author>Kobayashi, M ; Hanai, R</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p548-4801d9fbebf5a673969cfc83211dc2b761d8128c0285b9c88fd993c9f868bc0c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2001</creationdate><topic>Amino Acid Sequence</topic><topic>Base Sequence</topic><topic>Chromosomes</topic><topic>DNA Topoisomerases, Type I - genetics</topic><topic>DNA Topoisomerases, Type I - metabolism</topic><topic>Green Fluorescent Proteins</topic><topic>HeLa Cells</topic><topic>Humans</topic><topic>Isoenzymes - genetics</topic><topic>Isoenzymes - metabolism</topic><topic>Luminescent Proteins - metabolism</topic><topic>Mitosis - physiology</topic><topic>Molecular Sequence Data</topic><topic>Subcellular Fractions</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kobayashi, M</creatorcontrib><creatorcontrib>Hanai, R</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</collection><jtitle>Biochemical and biophysical research communications</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kobayashi, M</au><au>Hanai, R</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>M phase-specific association of human topoisomerase IIIbeta with chromosomes</atitle><jtitle>Biochemical and biophysical research communications</jtitle><addtitle>Biochem Biophys Res Commun</addtitle><date>2001-09-14</date><risdate>2001</risdate><volume>287</volume><issue>1</issue><spage>282</spage><epage>287</epage><pages>282-287</pages><issn>0006-291X</issn><abstract>Two isoforms, 1 and 2, of human DNA topoisomerase IIIbeta were expressed in HeLa cells as a fusion protein to the C-terminus of green fluorescent protein (GFP). The fusion protein of the isoform 1 was found to be localized to the nucleus, and to be associated with chromosomes during metaphase and anaphase. As yeast top3 mutants are known to exhibit phenotypes indicative of defective chromosome segregation, the result suggests that the isoform 1 of the human enzyme may also be involved in chromosome segregation. Two-hybrid screening for interaction partners of the isoform identified three candidate genes: CENP-F, a gene encoding a centromere protein and two genes of no known function, one of which was novel. The GFP fusion of the isoform 2 was found in the cytoplasm, indicating the nuclear localization signal sequence in the isoform 1 is in the C-terminal part that is different between the two isoforms.</abstract><cop>United States</cop><pmid>11549288</pmid><tpages>6</tpages></addata></record> |
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| ispartof | Biochemical and biophysical research communications, 2001-09, Vol.287 (1), p.282-287 |
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| subjects | Amino Acid Sequence Base Sequence Chromosomes DNA Topoisomerases, Type I - genetics DNA Topoisomerases, Type I - metabolism Green Fluorescent Proteins HeLa Cells Humans Isoenzymes - genetics Isoenzymes - metabolism Luminescent Proteins - metabolism Mitosis - physiology Molecular Sequence Data Subcellular Fractions |
| title | M phase-specific association of human topoisomerase IIIbeta with chromosomes |
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