2.50
Hdl Handle:
http://hdl.handle.net/10033/347315
Title:
An episomally replicating vector binds to the nuclear matrix protein SAF-A in vivo.
Authors:
Jenke, Bok Hee C; Fetzer, Christian P; Stehle, Isa M; Jönsson, Franziska; Fackelmayer, Frank O; Conradt, Harald; Bode, Jürgen; Lipps, Hans J
Abstract:
pEPI-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.
Affiliation:
Institute of Cell Biology, Stockumer Strasse 10, University of Witten/Herdecke, D-58448 Witten.
Citation:
An episomally replicating vector binds to the nuclear matrix protein SAF-A in vivo. 2002, 3 (4):349-54 EMBO Rep.
Journal:
EMBO reports
Issue Date:
Apr-2002
URI:
http://hdl.handle.net/10033/347315
DOI:
10.1093/embo-reports/kvf070
PubMed ID:
11897664
Type:
Article
Language:
en
ISSN:
1469-221X
Appears in Collections:
Publications of Dept. Gene Regulation and Differentiation (RDIF)

Full metadata record

DC FieldValue Language
dc.contributor.authorJenke, Bok Hee Cen
dc.contributor.authorFetzer, Christian Pen
dc.contributor.authorStehle, Isa Men
dc.contributor.authorJönsson, Franziskaen
dc.contributor.authorFackelmayer, Frank Oen
dc.contributor.authorConradt, Haralden
dc.contributor.authorBode, Jürgenen
dc.contributor.authorLipps, Hans Jen
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.en
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

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