含有共价DNA-蛋白质复合物的微小病毒（MVM）病毒粒子新群体的特征分析。

Characterization of novel populations of MVM virions containing covalent DNA-protein complexes.

作者信息

Faust E A, Brudzynska K, Morgan J

机构信息

Cancer Research Laboratory, University of Western Ontario, London, Canada.

出版信息

Virology. 1989 Jan;168(1):128-37. doi: 10.1016/0042-6822(89)90411-x.

DOI:10.1016/0042-6822(89)90411-x

PMID:2909986

Abstract

Virions of minute virus of mice were purified by sedimentation in sucrose gradients and chromatography on DEAE-cellulose columns and shown to consist of single-stranded viral DNA and the viral capsid polypeptides VP-1 (83 kDa) and VP-2 (64.5 kDa). A 63-kDa polypeptide distinct from the viral capsid polypeptide VP-3 (61.4 kDa) was found in some virion preparations. Virions sedimented at 135 and 110 S. The genomic single strands associated with purified 135 and 110 S virions were covalently bound to a protein as judged by the anomalous electrophoretic mobility of the DNA in agarose gels at pH 12.5. The protein was removed from the DNA by Pronase but remained bound after heating at 98 degrees in the presence of 0.1% sodium dodecyl sulfate. Nuclease digestion of the purified DNA-protein complex released several polypeptides ranging in size from 58 to 65 kDa. Restriction enzyme analysis of the purified DNA protein complex following its conversion to a duplex RF DNA in vitro showed that the protein was attached to the 5' termini of the DNA.

摘要

小鼠微小病毒的病毒粒子通过在蔗糖梯度中沉降和在DEAE - 纤维素柱上进行层析得以纯化，并显示其由单链病毒DNA以及病毒衣壳多肽VP - 1（83 kDa）和VP - 2（64.5 kDa）组成。在一些病毒粒子制剂中发现了一种与病毒衣壳多肽VP - 3（61.4 kDa）不同的63 kDa多肽。病毒粒子在135 S和110 S沉降。根据在pH 12.5的琼脂糖凝胶中DNA异常的电泳迁移率判断，与纯化的135 S和110 S病毒粒子相关的基因组单链与一种蛋白质共价结合。该蛋白质可被链霉蛋白酶从DNA上去除，但在0.1%十二烷基硫酸钠存在下于98℃加热后仍保持结合。对纯化的DNA - 蛋白质复合物进行核酸酶消化释放出几种大小在58至65 kDa之间的多肽。体外将纯化的DNA - 蛋白质复合物转化为双链RF DNA后进行的限制性内切酶分析表明，该蛋白质附着于DNA的5'末端。