An episomally replicating vector binds to the nuclear matrix protein SAF-A in vivo.
Jenke et al_final.pdf
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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.
The following license files are associated with this item:
- The scaffold/matrix attachment region binding protein hnRNP-U (SAF-A) is directly bound to chromosomal DNA in vivo: a chemical cross-linking study.
- Authors: Göhring F, Fackelmayer FO
- Issue date: 1997 Jul 8
- A vector based on the SV40 origin of replication and chromosomal S/MARs replicates episomally in CHO cells.
- Authors: Piechaczek C, Fetzer C, Baiker A, Bode J, Lipps HJ
- Issue date: 1999 Jan 15
- Nucleic-acid-binding properties of hnRNP-U/SAF-A, a nuclear-matrix protein which binds DNA and RNA in vivo and in vitro.
- Authors: Fackelmayer FO, Dahm K, Renz A, Ramsperger U, Richter A
- Issue date: 1994 Apr 15
- Nuclear scaffold/matrix attached region modules linked to a transcription unit are sufficient for replication and maintenance of a mammalian episome.
- Authors: Jenke AC, Stehle IM, Herrmann F, Eisenberger T, Baiker A, Bode J, Fackelmayer FO, Lipps HJ
- Issue date: 2004 Aug 3
- The novel SAR-binding domain of scaffold attachment factor A (SAF-A) is a target in apoptotic nuclear breakdown.
- Authors: Göhring F, Schwab BL, Nicotera P, Leist M, Fackelmayer FO
- Issue date: 1997 Dec 15