An episomally replicating vector binds to the nuclear matrix protein SAF-A in vivo.
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AuthorsJenke, Bok Hee C
Fetzer, Christian P
Stehle, Isa M
Fackelmayer, Frank O
Lipps, Hans J
MetadataShow full item record
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.
CitationAn episomally replicating vector binds to the nuclear matrix protein SAF-A in vivo. 2002, 3 (4):349-54 EMBO Rep.
AffiliationInstitute of Cell Biology, Stockumer Strasse 10, University of Witten/Herdecke, D-58448 Witten.
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- The scaffold/matrix attachment region binding protein hnRNP-U (SAF-A) is directly bound to chromosomal DNA in vivo: a chemical cross-linking study.
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