An episomally replicating vector binds to the nuclear matrix protein SAF-A in vivo.
dc.contributor.author | Jenke, Bok Hee C | |
dc.contributor.author | Fetzer, Christian P | |
dc.contributor.author | Stehle, Isa M | |
dc.contributor.author | Jönsson, Franziska | |
dc.contributor.author | Fackelmayer, Frank O | |
dc.contributor.author | Conradt, Harald | |
dc.contributor.author | Bode, Jürgen | |
dc.contributor.author | Lipps, Hans J | |
dc.date.accessioned | 2015-03-30T13:25:41Z | en |
dc.date.available | 2015-03-30T13:25:41Z | en |
dc.date.issued | 2002-04 | en |
dc.identifier.citation | An episomally replicating vector binds to the nuclear matrix protein SAF-A in vivo. 2002, 3 (4):349-54 EMBO Rep. | en |
dc.identifier.issn | 1469-221X | en |
dc.identifier.pmid | 11897664 | en |
dc.identifier.doi | 10.1093/embo-reports/kvf070 | en |
dc.identifier.uri | http://hdl.handle.net/10033/347315 | en |
dc.description.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. | |
dc.language.iso | en | en |
dc.subject.mesh | Animals | en |
dc.subject.mesh | Blotting, Southwestern | en |
dc.subject.mesh | Blotting, Western | en |
dc.subject.mesh | CHO Cells | en |
dc.subject.mesh | Cisplatin | en |
dc.subject.mesh | Cricetinae | en |
dc.subject.mesh | Genetic Vectors | en |
dc.subject.mesh | Heterogeneous-Nuclear Ribonucleoprotein U | en |
dc.subject.mesh | Heterogeneous-Nuclear Ribonucleoproteins | en |
dc.subject.mesh | Ribonucleoproteins | en |
dc.title | An episomally replicating vector binds to the nuclear matrix protein SAF-A in vivo. | en |
dc.type | Article | en |
dc.contributor.department | Institute of Cell Biology, Stockumer Strasse 10, University of Witten/Herdecke, D-58448 Witten. | en |
dc.identifier.journal | EMBO reports | en |
refterms.dateFOA | 2018-06-12T17:16:11Z | |
html.description.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. |