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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >DIFFERENTIAL REPLICATION OF A SINGLE, UV-INDUCED LESION IN THE LEADING OR LAGGING STRAND BY A HUMAN CELL EXTRACT - FORK UNCOUPLING OR GAP FORMATION
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DIFFERENTIAL REPLICATION OF A SINGLE, UV-INDUCED LESION IN THE LEADING OR LAGGING STRAND BY A HUMAN CELL EXTRACT - FORK UNCOUPLING OR GAP FORMATION

机译：人体提取物在前导或滞后链中单个，紫外线诱导的病变的差异复制-叉解偶联或间隙形成

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摘要

We have constructed simian virus 40 mini-replicons containing uniquely placed cis,syn-thymine dimers (T >$($) over bar T) for the analysis of leading- and lagging-strand bypass replication, Assaying for replication in a human cell-free extract through the analysis of full-size labeled product molecules and restriction fragments spanning the T >$($) over bar T site resulted in the following findings: (i) The primary site of synthesis blockage with T >$($) over bar T in either the leading or lagging strand was one nucleotide before the lesion, (ii) Replicative bypass of T >$($) over bar T was detected in both leading and lagging strands, The efficiency of synthesis past T >$($) over bar T was 22% for leading-strand T >$($) over bar T and 13% for lagging-strand T >$($) over bar T, (iii) The lagging-strand T >$($) over bar T resulted in blocked retrograde synthesis with the replication fork proceeding past the lesion, resulting in daughter molecules containing small gaps (form II' DNA), (iv) With T >$($) over bar T in the leading-strand template; both the leading and lagging strands were blocked, representing a stalled replication fork. Uncoupling of the concerted synthesis of the two strands of the replication fork was observed, resulting in preferential elongation of the undamaged lagging strand, These data support a model of selective reinitiation downstream from the lesion on lagging strands due to Okazaki synthesis, with no reinitiation close to the damage site on leading strands [Meneghini, R, & Hanawalt, P. C. (1976) Biochim. Biophys. Acta 425, 428-437].
机译：我们已经构建了猿猴病毒40个微型复制子，其中包含独特放置的顺式，顺式胸腺嘧啶二聚体（T条上方的T> $（$）），用于分析前导链和滞后链旁路复制，并用于在人细胞中复制通过分析在bar T位点上跨T> $（$）的全尺寸标记产物分子和限制性片段得到的游离提取物，得出以下发现：（i）T> $（$）的合成阻断的主要位点前导链或滞后链中的bar T是病变前一个核苷酸，（ii）在前导链和滞后链中均检测到T> $（$）超过bar T的复制旁路，合成效率超过T> $（$ ）在T条上，领先链T> $（$）在T条上为22％，在滞后链T> $（$）在T条上为（13），（iii）滞后T> $（$）超过T条导致逆转录合成受阻，复制叉越过病灶前进，导致子分子含有小缺口（II'DNA形式），（i v）在前导模板中，T之上的T> $（$）；前导链和滞后链均被阻塞，代表复制叉停滞。观察到复制叉的两条链的协调合成解偶联，导致未受损的落后链优先延伸。这些数据支持了由于冈崎合成而在滞后链上病变下游选择性重新初始化的模型，没有重新初始化关闭到前导链上的损伤位点[Meneghini，R，＆Hanawalt，PC（1976）Biochim。生物物理学。 Acta 425，428-437]。

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