Initiation of simian virus 40 DNA replication in vitro: aphidicolin causes accumulation of early-replicating intermediates and allows determination of the initial direction of DNA synthesis
Initiation of simian virus 40 DNA replication in vitro: aphidicolin causes accumulation of early-replicating intermediates and allows determination of the initial direction of DNA synthesis
复制标题
猿猴病毒 40 DNA 体外复制的启动：阿菲迪霉素导致早期复制中间体的积累，并允许确定 DNA 合成的初始方向
DOI：
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复制
发表时间：
1986
影响因子：
5.3
通讯作者：
M. DePamphilis
中科院分区：
文献类型：
--
作者：
R. Decker;M. Yamaguchi;R. Possenti;M. DePamphilis
Aphidicolin, a specific inhibitor of DNA polymerase alpha, provided a novel method for distinguishing between initiation of DNA synthesis at the simian virus 40 (SV40) origin of replication (ori) and continuation of replication beyond ori. In the presence of sufficient aphidicolin to inhibit total DNA synthesis by 50%, initiation of DNA replication in SV40 chromosomes or ori-containing plasmids continued in vitro, whereas DNA synthesis in the bulk of SV40 replicative intermediate DNA (RI) that had initiated replication in vivo was rapidly inhibited. This resulted in accumulation of early RI in which most nascent DNA was localized within a 600- to 700-base-pair region centered at ori. Accumulation of early RI was observed only under conditions that permitted initiation of SV40 ori-dependent, T-antigen-dependent DNA replication and only when aphidicolin was added to the in vitro system. Increasing aphidicolin concentrations revealed that DNA synthesis in the ori region was not completely resistant to aphidicolin but simply less sensitive than DNA synthesis at forks that were farther away. Since DNA synthesized in the presence of aphidicolin was concentrated in the 300 base pairs on the early gene side of ori, we conclude that the initial direction of DNA synthesis was the same as that of early mRNA synthesis, consistent with the model proposed by Hay and DePamphilis (Cell 28:767-779, 1982). The data were also consistent with initiation of the first DNA chains in ori by CV-1 cell DNA primase-DNA polymerase alpha. Synthesis of pppA/G(pN)6-8(pdN)21-23 chains on a single-stranded DNA template by a purified preparation of this enzyme was completely resistant to aphidicolin, and further incorporation of deoxynucleotide monophosphates was inhibited. Therefore, in the presence of aphidicolin, this enzyme could initiate RNA-primed DNA synthesis at ori first in the early gene direction and then in the late gene direction, but could not continue DNA synthesis for an extended distance.
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DOI：
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发表时间：
1984
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Gronostajski,RM;Field,J;Hurwitz,J
通讯作者：
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DOI：
--
发表时间：
1985
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Plevani,P;Foiani,M;Valsasnini,P;Badaracco,G;Cheriathundam,E;Chang,LM
通讯作者：
Chang,LM
启动猿猴病毒 40 DNA 复制所需因子的 DNA 结合位点。
DOI：
10.1073/pnas.83.6.1646
发表时间：
1986
影响因子：
11.1
作者：
Yamaguchi,M;DePamphilis,ML
通讯作者：
DePamphilis,ML
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影响因子：
5.6
作者：
Hay,RT;Hendrickson,EA;DePamphilis,ML
通讯作者：
DePamphilis,ML
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影响因子：
14.9
作者：
Armstrong,KA;Bauer,WR
通讯作者：
Bauer,WR