Miller B W, Williams J
Department of Biological Sciences, Carnegie-Mellon University, Pittsburgh, Pennsylvania 15213-3890.
J Virol. 1987 Nov;61(11):3630-4. doi: 10.1128/JVI.61.11.3630-3634.1987.
Early region 2B (E2B) of the group C adenoviruses encodes a number of proteins, including the 140-kilodalton DNA polymerase, which plays a role in the initiation of viral DNA replication. Temperature-sensitive (ts) mutants with mutations mapping to E2B are conditionally defective for both DNA replication in human cells and transformation of rat cells. Nucleotide sequence analysis shows that the E2B mutant ts36 possesses a single point mutation specific to the viral DNA polymerase; this transition of a C to a T at position 7623 changes leucine residue 249 in the polymerase to a phenylalanine. A wild-type (ts+) revertant possesses a codon specifying the original leucine at position 249. Phenotypic analysis of revertant and wild-type viruses derived by marker rescue from ts36 shows that these variants are wild type for both viral DNA replication and transformation. Thus, the single point mutation in the polymerase gene of ts36 is responsible for both defects.
C组腺病毒的早期区域2B(E2B)编码多种蛋白质,包括140千道尔顿的DNA聚合酶,该酶在病毒DNA复制起始过程中发挥作用。映射到E2B的温度敏感(ts)突变体在人类细胞中的DNA复制和大鼠细胞的转化方面均有条件缺陷。核苷酸序列分析表明,E2B突变体ts36在病毒DNA聚合酶上具有一个特定的单点突变;第7623位的C到T的转变将聚合酶中的亮氨酸残基249变为苯丙氨酸。野生型(ts+)回复体在第249位具有指定原始亮氨酸的密码子。通过从ts36进行标记拯救获得的回复体和野生型病毒的表型分析表明,这些变体在病毒DNA复制和转化方面均为野生型。因此,ts36聚合酶基因中的单点突变是这两种缺陷的原因。