Dornreiter I, Erdile L F, Gilbert I U, von Winkler D, Kelly T J, Fanning E
Institute for Biochemistry, Munich, Germany.
EMBO J. 1992 Feb;11(2):769-76. doi: 10.1002/j.1460-2075.1992.tb05110.x.
The purified human single-stranded DNA binding protein, replication protein A (RP-A), forms specific complexes with purified SV40 large T antigen and with purified DNA polymerase alpha-primase, as shown by ELISA and a modified immunoblotting technique. RP-A associated efficiently with the isolated primase, as well as with intact polymerase alpha-primase. The 70 kDa subunit of RP-A was sufficient for association with polymerase alpha-primase. Purified SV40 large T antigen bound to intact RP-A and to polymerase-primase, but not to any of the separated subunits of RP-A or to the isolated primase. These results suggest that the specific protein-protein interactions between RP-A, polymerase-primase and T antigen may play a role in the initiating of SV40 DNA replication.
纯化的人单链DNA结合蛋白，即复制蛋白A（RP-A），与纯化的SV40大T抗原以及纯化的DNA聚合酶α-引发酶形成特异性复合物，这通过酶联免疫吸附测定（ELISA）和改良的免疫印迹技术得以证明。RP-A能有效地与分离出的引发酶以及完整的聚合酶α-引发酶结合。RP-A的70 kDa亚基足以与聚合酶α-引发酶结合。纯化的SV40大T抗原与完整的RP-A以及聚合酶-引发酶结合，但不与RP-A的任何分离亚基或分离出的引发酶结合。这些结果表明，RP-A、聚合酶-引发酶和T抗原之间的特异性蛋白质-蛋白质相互作用可能在SV40 DNA复制的起始过程中发挥作用。
