Nuclear Clusterin/XIP8, an X-Ray-Induced Ku70-Binding Protein That Signals Cell Death
Clusterin [CLU, a.k.a. TRPM-2, SGP-2, or ionizing radiation (IR)-induced protein-8(XIP8)] was implicated in apoptosis, tissue injury, and aging. Its function remains elusive. We reisolated CLU/XIP8 by yeast two-hybrid analyses using as bait the DNA double-strand break repair protein Ku70. We show th...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 2000-05, Vol.97 (11), p.5907-5912 |
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| container_title | Proceedings of the National Academy of Sciences - PNAS |
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| creator | Yang, Chin-Rang Leskov, Konstantin Hosley-Eberlein, Kelly Criswell, Tracy Pink, John J. Kinsella, Timothy J. Boothman, David A. |
| description | Clusterin [CLU, a.k.a. TRPM-2, SGP-2, or ionizing radiation (IR)-induced protein-8(XIP8)] was implicated in apoptosis, tissue injury, and aging. Its function remains elusive. We reisolated CLU/XIP8 by yeast two-hybrid analyses using as bait the DNA double-strand break repair protein Ku70. We show that a delayed (2-3 days), low-dose (0.02-10 Gy) IR-inducible nuclear CLU/XIP8 protein coimmunoprecipitated and colocalized (by confocal microscopy) in vivo with Ku70/Ku80, a DNA damage sensor and key double-strand break repair protein, in human MCF-7:WS8 breast cancer cells. Overexpression of nuclear CLU/XIP8 or its minimal Ku70 binding domain (120 aa of CLU/XIP8 C terminus) in nonirradiated MCF-7:WS8 cells dramatically reduced cell growth and colony-forming ability concomitant with increased G1cell cycle check-point arrest and increased cell death. Enhanced expression and accumulation of nuclear CLU/XIP8-Ku70/Ku80 complexes appears to be an important cell death signal after IR exposure. |
| doi_str_mv | 10.1073/pnas.97.11.5907 |
| format | Article |
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TRPM-2, SGP-2, or ionizing radiation (IR)-induced protein-8(XIP8)] was implicated in apoptosis, tissue injury, and aging. Its function remains elusive. We reisolated CLU/XIP8 by yeast two-hybrid analyses using as bait the DNA double-strand break repair protein Ku70. We show that a delayed (2-3 days), low-dose (0.02-10 Gy) IR-inducible nuclear CLU/XIP8 protein coimmunoprecipitated and colocalized (by confocal microscopy) in vivo with Ku70/Ku80, a DNA damage sensor and key double-strand break repair protein, in human MCF-7:WS8 breast cancer cells. Overexpression of nuclear CLU/XIP8 or its minimal Ku70 binding domain (120 aa of CLU/XIP8 C terminus) in nonirradiated MCF-7:WS8 cells dramatically reduced cell growth and colony-forming ability concomitant with increased G1cell cycle check-point arrest and increased cell death. Enhanced expression and accumulation of nuclear CLU/XIP8-Ku70/Ku80 complexes appears to be an important cell death signal after IR exposure.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.97.11.5907</identifier><identifier>PMID: 10823943</identifier><language>eng</language><publisher>United States: National Academy of Sciences of the United States of America</publisher><subject>Adenocarcinoma - pathology ; Amino Acid Motifs ; Antibodies ; Antigens, Nuclear ; Biological Sciences ; Breast Neoplasms - pathology ; Cell cycle ; Cell death ; Cell Death - physiology ; Cell growth ; Cell nucleus ; Cell Nucleus - metabolism ; Cellular biology ; Clusterin ; Complementary DNA ; Deoxyribonucleic acid ; DNA ; DNA Damage ; DNA Helicases ; DNA Repair - physiology ; DNA, Complementary - genetics ; DNA, Neoplasm - radiation effects ; DNA-Activated Protein Kinase ; DNA-Binding Proteins - metabolism ; Female ; Flow Cytometry ; Gene Expression Regulation, Neoplastic - radiation effects ; Gene Library ; Genes, Reporter ; Glycoproteins - chemistry ; Glycoproteins - genetics ; Glycoproteins - physiology ; Green Fluorescent Proteins ; Humans ; Infrared radiation ; Ku Autoantigen ; Ku70-binding protein ; Ku80 antigen ; Luminescent Proteins - genetics ; Microscopy, Confocal ; Molecular Chaperones ; Nuclear Proteins - metabolism ; Protein Binding ; Protein Precursors - metabolism ; Protein-Serine-Threonine Kinases - physiology ; Proteins ; Saccharomyces cerevisiae - genetics ; Saccharomyces cerevisiae Proteins ; Tumor Cells, Cultured ; Two-Hybrid System Techniques ; XIP8 protein ; Yeasts</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 2000-05, Vol.97 (11), p.5907-5912</ispartof><rights>Copyright 1993-2000 National Academy of Sciences of the United States of America</rights><rights>Copyright National Academy of Sciences May 23, 2000</rights><rights>Copyright © 2000, The National Academy of Sciences 2000</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c566t-a923a8ab6a7083ea1d8900171256af94a7997cd5dac5464ba3a67ba0903574ed3</citedby><cites>FETCH-LOGICAL-c566t-a923a8ab6a7083ea1d8900171256af94a7997cd5dac5464ba3a67ba0903574ed3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/97/11.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/122555$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/122555$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,723,776,780,799,881,27901,27902,53766,53768,57992,58225</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10823943$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Yang, Chin-Rang</creatorcontrib><creatorcontrib>Leskov, Konstantin</creatorcontrib><creatorcontrib>Hosley-Eberlein, Kelly</creatorcontrib><creatorcontrib>Criswell, Tracy</creatorcontrib><creatorcontrib>Pink, John J.</creatorcontrib><creatorcontrib>Kinsella, Timothy J.</creatorcontrib><creatorcontrib>Boothman, David A.</creatorcontrib><title>Nuclear Clusterin/XIP8, an X-Ray-Induced Ku70-Binding Protein That Signals Cell Death</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>Clusterin [CLU, a.k.a. TRPM-2, SGP-2, or ionizing radiation (IR)-induced protein-8(XIP8)] was implicated in apoptosis, tissue injury, and aging. Its function remains elusive. We reisolated CLU/XIP8 by yeast two-hybrid analyses using as bait the DNA double-strand break repair protein Ku70. We show that a delayed (2-3 days), low-dose (0.02-10 Gy) IR-inducible nuclear CLU/XIP8 protein coimmunoprecipitated and colocalized (by confocal microscopy) in vivo with Ku70/Ku80, a DNA damage sensor and key double-strand break repair protein, in human MCF-7:WS8 breast cancer cells. Overexpression of nuclear CLU/XIP8 or its minimal Ku70 binding domain (120 aa of CLU/XIP8 C terminus) in nonirradiated MCF-7:WS8 cells dramatically reduced cell growth and colony-forming ability concomitant with increased G1cell cycle check-point arrest and increased cell death. Enhanced expression and accumulation of nuclear CLU/XIP8-Ku70/Ku80 complexes appears to be an important cell death signal after IR exposure.</description><subject>Adenocarcinoma - pathology</subject><subject>Amino Acid Motifs</subject><subject>Antibodies</subject><subject>Antigens, Nuclear</subject><subject>Biological Sciences</subject><subject>Breast Neoplasms - pathology</subject><subject>Cell cycle</subject><subject>Cell death</subject><subject>Cell Death - physiology</subject><subject>Cell growth</subject><subject>Cell nucleus</subject><subject>Cell Nucleus - metabolism</subject><subject>Cellular biology</subject><subject>Clusterin</subject><subject>Complementary DNA</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA Damage</subject><subject>DNA Helicases</subject><subject>DNA Repair - physiology</subject><subject>DNA, Complementary - genetics</subject><subject>DNA, Neoplasm - radiation effects</subject><subject>DNA-Activated Protein Kinase</subject><subject>DNA-Binding Proteins - metabolism</subject><subject>Female</subject><subject>Flow Cytometry</subject><subject>Gene Expression Regulation, Neoplastic - radiation effects</subject><subject>Gene Library</subject><subject>Genes, Reporter</subject><subject>Glycoproteins - chemistry</subject><subject>Glycoproteins - genetics</subject><subject>Glycoproteins - physiology</subject><subject>Green Fluorescent Proteins</subject><subject>Humans</subject><subject>Infrared radiation</subject><subject>Ku Autoantigen</subject><subject>Ku70-binding protein</subject><subject>Ku80 antigen</subject><subject>Luminescent Proteins - genetics</subject><subject>Microscopy, Confocal</subject><subject>Molecular Chaperones</subject><subject>Nuclear Proteins - metabolism</subject><subject>Protein Binding</subject><subject>Protein Precursors - metabolism</subject><subject>Protein-Serine-Threonine Kinases - physiology</subject><subject>Proteins</subject><subject>Saccharomyces cerevisiae - genetics</subject><subject>Saccharomyces cerevisiae Proteins</subject><subject>Tumor Cells, Cultured</subject><subject>Two-Hybrid System Techniques</subject><subject>XIP8 protein</subject><subject>Yeasts</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU1vEzEQhi0EomnhzAEJrTiUSzcZ22t7LfXSBgoRFVTQSr1Zk10n2WjjDf5A9N-zqxRIOcBppJnnna-XkBcUxhQUn2wdhrFWY0rHQoN6REYUNM1loeExGQEwlZcFKw7IYQhrANCihKfkgELJuC74iNx8SlVr0WfTNoVofeMmt7Or8iRDl93mX_Aun7k6VbbOPiYF-Xnj6sYtsyvfRdu47HqFMfvaLB22IZvats3eWoyrZ-TJos_Y5_fxiNxcvLuefsgvP7-fTc8u80pIGXPUjGOJc4kKSm6R1qUGoIoyIXGhC1Raq6oWNVaikMUcOUo1R9DAhSpszY_I6a7vNs03tq6six5bs_XNBv2d6bAxDyuuWZll993QUnDWy4_v5b77lmyIZtOEqr8Cne1SMIr2m7BS_RekSrBCStmDr_8C113yw3cMA8oVl5z30GQHVb4LwdvF74UpmMFWM9hqtDKUmsHWXvFq_849fufjHjAof5UfdHjzT8AsUttG-yP25MsduQ6x839GMSaE4D8BrCa9rw</recordid><startdate>20000523</startdate><enddate>20000523</enddate><creator>Yang, Chin-Rang</creator><creator>Leskov, Konstantin</creator><creator>Hosley-Eberlein, Kelly</creator><creator>Criswell, Tracy</creator><creator>Pink, John J.</creator><creator>Kinsella, Timothy J.</creator><creator>Boothman, David A.</creator><general>National Academy of Sciences of the United States of America</general><general>National Acad Sciences</general><general>National Academy of Sciences</general><general>The National Academy of Sciences</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>7QG</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TK</scope><scope>7TM</scope><scope>7TO</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20000523</creationdate><title>Nuclear Clusterin/XIP8, an X-Ray-Induced Ku70-Binding Protein That Signals Cell Death</title><author>Yang, Chin-Rang ; Leskov, Konstantin ; Hosley-Eberlein, Kelly ; Criswell, Tracy ; Pink, John J. ; Kinsella, Timothy J. ; Boothman, David A.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c566t-a923a8ab6a7083ea1d8900171256af94a7997cd5dac5464ba3a67ba0903574ed3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Adenocarcinoma - pathology</topic><topic>Amino Acid Motifs</topic><topic>Antibodies</topic><topic>Antigens, Nuclear</topic><topic>Biological Sciences</topic><topic>Breast Neoplasms - pathology</topic><topic>Cell cycle</topic><topic>Cell death</topic><topic>Cell Death - physiology</topic><topic>Cell growth</topic><topic>Cell nucleus</topic><topic>Cell Nucleus - metabolism</topic><topic>Cellular biology</topic><topic>Clusterin</topic><topic>Complementary DNA</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>DNA Damage</topic><topic>DNA Helicases</topic><topic>DNA Repair - physiology</topic><topic>DNA, Complementary - genetics</topic><topic>DNA, Neoplasm - radiation effects</topic><topic>DNA-Activated Protein Kinase</topic><topic>DNA-Binding Proteins - metabolism</topic><topic>Female</topic><topic>Flow Cytometry</topic><topic>Gene Expression Regulation, Neoplastic - radiation effects</topic><topic>Gene Library</topic><topic>Genes, Reporter</topic><topic>Glycoproteins - chemistry</topic><topic>Glycoproteins - genetics</topic><topic>Glycoproteins - physiology</topic><topic>Green Fluorescent Proteins</topic><topic>Humans</topic><topic>Infrared radiation</topic><topic>Ku Autoantigen</topic><topic>Ku70-binding protein</topic><topic>Ku80 antigen</topic><topic>Luminescent Proteins - genetics</topic><topic>Microscopy, Confocal</topic><topic>Molecular Chaperones</topic><topic>Nuclear Proteins - metabolism</topic><topic>Protein Binding</topic><topic>Protein Precursors - metabolism</topic><topic>Protein-Serine-Threonine Kinases - physiology</topic><topic>Proteins</topic><topic>Saccharomyces cerevisiae - genetics</topic><topic>Saccharomyces cerevisiae Proteins</topic><topic>Tumor Cells, Cultured</topic><topic>Two-Hybrid System Techniques</topic><topic>XIP8 protein</topic><topic>Yeasts</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Yang, Chin-Rang</creatorcontrib><creatorcontrib>Leskov, Konstantin</creatorcontrib><creatorcontrib>Hosley-Eberlein, Kelly</creatorcontrib><creatorcontrib>Criswell, Tracy</creatorcontrib><creatorcontrib>Pink, John J.</creatorcontrib><creatorcontrib>Kinsella, Timothy J.</creatorcontrib><creatorcontrib>Boothman, David A.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Oncogenes and Growth Factors Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Yang, Chin-Rang</au><au>Leskov, Konstantin</au><au>Hosley-Eberlein, Kelly</au><au>Criswell, Tracy</au><au>Pink, John J.</au><au>Kinsella, Timothy J.</au><au>Boothman, David A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Nuclear Clusterin/XIP8, an X-Ray-Induced Ku70-Binding Protein That Signals Cell Death</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>2000-05-23</date><risdate>2000</risdate><volume>97</volume><issue>11</issue><spage>5907</spage><epage>5912</epage><pages>5907-5912</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>Clusterin [CLU, a.k.a. TRPM-2, SGP-2, or ionizing radiation (IR)-induced protein-8(XIP8)] was implicated in apoptosis, tissue injury, and aging. Its function remains elusive. We reisolated CLU/XIP8 by yeast two-hybrid analyses using as bait the DNA double-strand break repair protein Ku70. We show that a delayed (2-3 days), low-dose (0.02-10 Gy) IR-inducible nuclear CLU/XIP8 protein coimmunoprecipitated and colocalized (by confocal microscopy) in vivo with Ku70/Ku80, a DNA damage sensor and key double-strand break repair protein, in human MCF-7:WS8 breast cancer cells. Overexpression of nuclear CLU/XIP8 or its minimal Ku70 binding domain (120 aa of CLU/XIP8 C terminus) in nonirradiated MCF-7:WS8 cells dramatically reduced cell growth and colony-forming ability concomitant with increased G1cell cycle check-point arrest and increased cell death. Enhanced expression and accumulation of nuclear CLU/XIP8-Ku70/Ku80 complexes appears to be an important cell death signal after IR exposure.</abstract><cop>United States</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>10823943</pmid><doi>10.1073/pnas.97.11.5907</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Adenocarcinoma - pathology Amino Acid Motifs Antibodies Antigens, Nuclear Biological Sciences Breast Neoplasms - pathology Cell cycle Cell death Cell Death - physiology Cell growth Cell nucleus Cell Nucleus - metabolism Cellular biology Clusterin Complementary DNA Deoxyribonucleic acid DNA DNA Damage DNA Helicases DNA Repair - physiology DNA, Complementary - genetics DNA, Neoplasm - radiation effects DNA-Activated Protein Kinase DNA-Binding Proteins - metabolism Female Flow Cytometry Gene Expression Regulation, Neoplastic - radiation effects Gene Library Genes, Reporter Glycoproteins - chemistry Glycoproteins - genetics Glycoproteins - physiology Green Fluorescent Proteins Humans Infrared radiation Ku Autoantigen Ku70-binding protein Ku80 antigen Luminescent Proteins - genetics Microscopy, Confocal Molecular Chaperones Nuclear Proteins - metabolism Protein Binding Protein Precursors - metabolism Protein-Serine-Threonine Kinases - physiology Proteins Saccharomyces cerevisiae - genetics Saccharomyces cerevisiae Proteins Tumor Cells, Cultured Two-Hybrid System Techniques XIP8 protein Yeasts |
| title | Nuclear Clusterin/XIP8, an X-Ray-Induced Ku70-Binding Protein That Signals Cell Death |
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