Deletion of the E4 region of the genome produces adenovirus DNA concatemers.
Deletion of the E4 region of the genome produces adenovirus DNA concatemers.
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基因组 E4 区域的缺失会产生腺病毒 DNA 多联体。
DOI：
10.1073/pnas.91.1.153
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发表时间：
1994
影响因子：
11.1
通讯作者：
Ginsberg,HS
中科院分区：
文献类型：
--
作者：
Weiden,MD;Ginsberg,HS
使用在腺病毒基因组的E4区中含有大缺失的两个突变体H5 dl 366（91.9-98.3图单位）和H2 dl 808（93.0-97.1图单位）来研究E4基因在腺病毒DNA合成中的作用。感染KB人表皮样癌细胞与任一突变体导致生产的大串联体的病毒DNA。然而，当突变体感染W162细胞时，仅产生单体病毒基因组形式，W162细胞是用E4基因转化并表达E4基因的猴肾细胞系。因此，可扩散的E4基因产物补充了E4突变体表型。在dl 366和dl 808感染的KB细胞中产生的病毒DNA多联体没有任何特定的单体连接方向：接头由头对头、头对尾和尾对尾接头组成。连接点是共价连接的分子，但分子并没有精确连接，限制性内切酶图谱显示连接片段的大小分布不均匀。还研究了一系列破坏单个E4开放阅读框（ORF）的突变体：没有一个显示出与dl 366或dl 808相似的表型。然而，在ORF 3和ORF 6中含有缺陷的突变体表现出多联体表型，表明阻止多联体形成的基因冗余。这些数据表明，E4 ORF 3和6表达的功能，调节病毒DNA复制的关键和多联体中间体可能存在于腺病毒DNA合成。
Two mutants containing large deletions in the E4 region of the adenovirus genome H5dl366 (91.9-98.3 map units) and H2dl808 (93.0-97.1 map units) were used to investigate the role of E4 genes in adenovirus DNA synthesis. Infection of KB human epidermoid carcinoma cells with either mutant resulted in production of large concatemers of viral DNA. Only monomer viral genome forms were produced, however, when mutants infected W162 cells, a monkey kidney cell line transformed with and expressing the E4 genes. Diffusible E4 gene products, therefore, complement the E4 mutant phenotype. The viral DNA concatemers produced in dl366- and dl808-infected KB cells did not have any specific orientation of monomer joining: the junctions consisted of head-to-head, head-to-tail, and tail-to-tail joints. The junctions were covalently linked molecules, but molecules were not precisely joined, and restriction enzyme maps revealed a heterogeneous size distribution of junction fragments. A series of mutants that disrupted single E4 open reading frames (ORFs) was also studied: none showed phenotypes similar to that of dl366 or dl808. Mutants containing defects in both ORF3 and ORF6, however, manifested the concatemer phenotype, indicating redundancy in genes preventing concatemer formation. These data suggest that the E4 ORFs 3 and 6 express functions critical for regulation of viral DNA replication and that concatemer intermediates may exist during adenovirus DNA synthesis.