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dc.contributor.authorJenke, Bok Hee C
dc.contributor.authorFetzer, Christian P
dc.contributor.authorStehle, Isa M
dc.contributor.authorJönsson, Franziska
dc.contributor.authorFackelmayer, Frank O
dc.contributor.authorConradt, Harald
dc.contributor.authorBode, Jürgen
dc.contributor.authorLipps, Hans J
dc.date.accessioned2015-03-30T13:25:41Zen
dc.date.available2015-03-30T13:25:41Zen
dc.date.issued2002-04en
dc.identifier.citationAn episomally replicating vector binds to the nuclear matrix protein SAF-A in vivo. 2002, 3 (4):349-54 EMBO Rep.en
dc.identifier.issn1469-221Xen
dc.identifier.pmid11897664en
dc.identifier.doi10.1093/embo-reports/kvf070en
dc.identifier.urihttp://hdl.handle.net/10033/347315en
dc.description.abstractpEPI-1, a vector in which a chromosomal scaffold/matrix-attached region (S/MAR) is linked to the simian virus 40 origin of replication, is propagated episomally in CHO cells in the absence of the virally encoded large T-antigen and is stably maintained in the absence of selection pressure. It has been suggested that mitotic stability is provided by a specific interaction of this vector with components of the nuclear matrix. We studied the interactions of pEPI-1 by crosslinking with cis-diamminedichloroplatinum II, after which it is found to copurify with the nuclear matrix. In a south-western analysis, the vector shows exclusive binding to hnRNP-U/SAF-A, a multifunctional scaffold/matrix specific factor. Immunoprecipitation of the crosslinked DNA-protein complex demonstrates that pEPI-1 is bound to this protein in vivo. These data provide the first experimental evidence for the binding of an artificial episome to a nuclear matrix protein in vivo and the basis for understanding the mitotic stability of this novel vector class.
dc.language.isoenen
dc.subject.meshAnimalsen
dc.subject.meshBlotting, Southwesternen
dc.subject.meshBlotting, Westernen
dc.subject.meshCHO Cellsen
dc.subject.meshCisplatinen
dc.subject.meshCricetinaeen
dc.subject.meshGenetic Vectorsen
dc.subject.meshHeterogeneous-Nuclear Ribonucleoprotein Uen
dc.subject.meshHeterogeneous-Nuclear Ribonucleoproteinsen
dc.subject.meshRibonucleoproteinsen
dc.titleAn episomally replicating vector binds to the nuclear matrix protein SAF-A in vivo.en
dc.typeArticleen
dc.contributor.departmentInstitute of Cell Biology, Stockumer Strasse 10, University of Witten/Herdecke, D-58448 Witten.en
dc.identifier.journalEMBO reportsen
refterms.dateFOA2018-06-12T17:16:11Z
html.description.abstractpEPI-1, a vector in which a chromosomal scaffold/matrix-attached region (S/MAR) is linked to the simian virus 40 origin of replication, is propagated episomally in CHO cells in the absence of the virally encoded large T-antigen and is stably maintained in the absence of selection pressure. It has been suggested that mitotic stability is provided by a specific interaction of this vector with components of the nuclear matrix. We studied the interactions of pEPI-1 by crosslinking with cis-diamminedichloroplatinum II, after which it is found to copurify with the nuclear matrix. In a south-western analysis, the vector shows exclusive binding to hnRNP-U/SAF-A, a multifunctional scaffold/matrix specific factor. Immunoprecipitation of the crosslinked DNA-protein complex demonstrates that pEPI-1 is bound to this protein in vivo. These data provide the first experimental evidence for the binding of an artificial episome to a nuclear matrix protein in vivo and the basis for understanding the mitotic stability of this novel vector class.


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