一种游离型复制载体在体内与核基质蛋白SAF-A结合。
An episomally replicating vector binds to the nuclear matrix protein SAF-A in vivo.
作者信息
Jenke Bok Hee C, Fetzer Christian P, Stehle Isa M, Jönsson Franziska, Fackelmayer Frank O, Conradt Harald, Bode Jürgen, Lipps Hans J
机构信息
Institute of Cell Biology, Stockumer Strasse 10, University of Witten/Herdecke, D-58448 Witten, 1Heinrich Pette-Institute, University of Hamburg, Hamburg, Germany.
出版信息
EMBO Rep. 2002 Apr;3(4):349-54. doi: 10.1093/embo-reports/kvf070. Epub 2002 Mar 15.
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.
pEPI-1是一种载体，其中染色体支架/基质附着区域（S/MAR）与猿猴病毒40复制起点相连，在没有病毒编码的大T抗原的情况下，它在CHO细胞中以附加体形式繁殖，并且在没有选择压力的情况下能稳定维持。有人提出，这种载体与核基质成分的特异性相互作用提供了有丝分裂稳定性。我们通过与顺二氨二氯铂（II）交联来研究pEPI-1的相互作用，之后发现它与核基质共纯化。在西南分析中，该载体显示出与hnRNP-U/SAF-A（一种多功能支架/基质特异性因子）的特异性结合。对交联的DNA-蛋白质复合物进行免疫沉淀表明，pEPI-1在体内与这种蛋白质结合。这些数据为人工附加体在体内与核基质蛋白结合提供了首个实验证据，也为理解这类新型载体的有丝分裂稳定性奠定了基础。
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