Mirza A
Institut für Molekularbiologie (Tumorforschung), Universitätsklinikum Essen, Federal Republic of Germany.
Arch Virol. 1988;102(3-4):245-62. doi: 10.1007/BF01310829.
Adenovirus type 12 transcriptional complexes were isolated from cells during the early phase of infection. Sedimentation analysis identified a fast sedimenting complex type I and a slow sedimenting complex type II. Both complexes made virus specific RNA complementary to all the early genes and both contained viral DNA, which in type II but not in type I had nucleosome like configuration. Analysis of the proteins of the complexes with antiserum against Ad 12 EIa-beta-galactosidase fusion protein expressed in E. coli demonstrated the following: (a) type I complex contained EIa 45 K protein, which co-precipitated with cellular proteins of mol. wt. 42, 58, and 60 K, (b) type II complex contained EIa 47 K protein, which co-precipitated with major cellular proteins of 35, 40-46 K and minor proteins of 58, 60, 68, 76, 86, and 120-150 K. Association of EIa specific and cellular proteins to transcriptional complexes was sensitive to both 1 M NaCl and DNAse I indicating the DNA binding nature of these proteins. Treatment of transcriptional complexes with 1 M NaCl or DNase I released EIa proteins, which still remained strongly bound to cellular proteins. These findings suggested that EIa proteins bind to viral DNA and that this binding is probably mediated by cellular proteins.
在感染早期从细胞中分离出12型腺病毒转录复合物。沉降分析鉴定出一种快速沉降的I型复合物和一种缓慢沉降的II型复合物。两种复合物都产生与所有早期基因互补的病毒特异性RNA，并且都含有病毒DNA，其中II型复合物而非I型复合物具有核小体样结构。用针对在大肠杆菌中表达的Ad 12 EIa-β-半乳糖苷酶融合蛋白的抗血清分析复合物的蛋白质，结果如下：（a）I型复合物含有EIa 45K蛋白，它与分子量为42K、58K和60K的细胞蛋白共沉淀；（b）II型复合物含有EIa 47K蛋白，它与35K、40 - 46K的主要细胞蛋白以及58K、60K、68K、76K、86K和120 - 150K的次要蛋白共沉淀。EIa特异性蛋白和细胞蛋白与转录复合物的结合对1M NaCl和DNA酶I均敏感，表明这些蛋白具有DNA结合特性。用1M NaCl或DNA酶I处理转录复合物会释放出EIa蛋白，这些蛋白仍与细胞蛋白紧密结合。这些发现表明EIa蛋白与病毒DNA结合，并且这种结合可能由细胞蛋白介导。
