Induction of apoptosis by overexpression of the DNA-binding and DNA-PK-activating protein C1D

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dc.citation.epage2232en_US
dc.citation.issueNumber13en_US
dc.citation.spage2223en_US
dc.citation.volumeNumber112en_US
dc.contributor.authorRothbarth, K.en_US
dc.contributor.authorSpiess, E.en_US
dc.contributor.authorJuodka, B.en_US
dc.contributor.authorYavuzer, U.en_US
dc.contributor.authorNehls, P.en_US
dc.contributor.authorStammer, H.en_US
dc.contributor.authorWerner, D.en_US
dc.date.accessioned2016-02-08T10:41:08Z
dc.date.available2016-02-08T10:41:08Z
dc.date.issued1999en_US
dc.departmentDepartment of Molecular Biology and Geneticsen_US
dc.description.abstractApoptosis is induced in various tumor cell lines by vector-dependent overexpression of the conserved gene C1D that encodes a DNA-binding and DNA-PK-activating protein. C1D is physiologically expressed in 50 human tissues tested, which points to its basic cellular function. The expression of this gene must be tightly regulated because elevated levels of C1D protein, e.g. those induced by transient vector-dependent expression, result in apoptotic cell death. Cells transfected with C1D-expressing constructs show terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling of DNA ends. Transfections with constructs in which C1D is expressed in fusion with the (enhanced) green fluorescent protein from A. victoria (EGFP) allow the transfected cells to be identified and the morphological changes induced to be traced. Starting from intense nuclear spots, green fluorescence reflecting C1D expression increases dramatically at 12-24 hours post-transfection. Expression of C1D-EGFP protein is accompanied by morphological changes typical of apoptotic cell death, e.g. cytoplasmic vacuolation, membrane blebbing and nuclear disintegration. Cell shrinkage and detachment from extracellular matrix are observed in monolayer cultures while suspension cells become progressively flattened. The facility to differentiate between transfected and non-transfected cells reveals that non-transfected cells co-cultured with transfected cells also show the morphological changes of apoptosis, which points to a bystander effect. C1D-dependent apoptosis is not induced in cells with non-functional p53. Accordingly, C1D-induced apoptosis is discussed in relation to its potential to activate DNA-PK, which has been considered to act as an upstream activator of p53.en_US
dc.description.provenanceMade available in DSpace on 2016-02-08T10:41:08Z (GMT). No. of bitstreams: 1 bilkent-research-paper.pdf: 70227 bytes, checksum: 26e812c6f5156f83f0e77b261a471b5a (MD5) Previous issue date: 1999en
dc.identifier.issn0021-9533
dc.identifier.urihttp://hdl.handle.net/11693/25220
dc.language.isoEnglishen_US
dc.source.titleJournal of Cell Scienceen_US
dc.subjectApoptosisen_US
dc.subjectBystander effecten_US
dc.subjectC1D geneen_US
dc.subjectDNA-binding proteinen_US
dc.subjectDNA-PKen_US
dc.subjectEnhanced green fluorescent proteinen_US
dc.subjectVector-dependent overexpressionen_US
dc.subjectDNA binding proteinen_US
dc.subjectDNA dependent protein kinaseen_US
dc.subjectDNA nucleotidylexotransferaseen_US
dc.titleInduction of apoptosis by overexpression of the DNA-binding and DNA-PK-activating protein C1Den_US
dc.typeArticleen_US

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