Miller M R, Ulrich R G, Wang T S, Korn D
J Biol Chem. 1985 Jan 10;260(1):134-8.
Monoclonal neutralizing antibodies against DNA polymerase-alpha substantially inhibit nuclear DNA replication in lysolecithin-permeabilized cultured human fibroblasts. The degree of inhibition of DNA synthesis is proportional to antibody concentration, and the effect is specific in that RNA synthesis measured under the same experimental conditions is unperturbed. Autoradiographic data demonstrate that the magnitude of the inhibition measured in the mass culture reflects the uniform response of all the constituent cells in the target population. These observations confirm the participation of DNA polymerase-alpha in replicative DNA synthesis and identify a versatile, novel approach to the dissection of mammalian processes of DNA replication and repair.
针对DNA聚合酶α的单克隆中和抗体可显著抑制溶血卵磷脂通透的培养人成纤维细胞中的核DNA复制。DNA合成的抑制程度与抗体浓度成正比,并且这种效应具有特异性,因为在相同实验条件下测得的RNA合成不受干扰。放射自显影数据表明,在大规模培养中测得的抑制程度反映了靶细胞群体中所有组成细胞的一致反应。这些观察结果证实了DNA聚合酶α参与复制性DNA合成,并确定了一种通用的、新颖的方法来剖析哺乳动物的DNA复制和修复过程。