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大肠杆菌微型染色体在oriC和蛋白质n'识别位点处的复制起始。体外启动DNA合成的两种模式。

Initiation of Escherichia coli minichromosome replication at oriC and at protein n' recognition sites. Two modes for initiating DNA synthesis in vitro.

作者信息

Seufert W, Messer W

出版信息

EMBO J. 1986 Dec 1;5(12):3401-6. doi: 10.1002/j.1460-2075.1986.tb04656.x.

DOI:10.1002/j.1460-2075.1986.tb04656.x
PMID:3028783
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1167339/
Abstract

The start sites for leading and lagging DNA strands were determined in vitro with minichromosomes as templates. Fragments from replication intermediates were analyzed by hybridization to single-stranded probes. Leading strand synthesis in the counterclockwise direction was found to originate in or close to (position 248 to -44) the minimal origin. Complementary lagging strand synthesis started several positions to the left outside of oriC. The results suggest in addition a concerted synthesis of leading and lagging strands following the dnaA directed assembly of initiation proteins at double-stranded oricC DNA (pre-replisome). In addition, DNA synthesis could initiate at protein n' recognition sequences located within and clockwise to the asnA gene. Initiation at n' sites was dependent on protein i activity, whereas leading and lagging strand initiation in the oriC region was not affected by protein i. Our results argue against an involvement of the phi X174-type primosome in the initiation of discontinuous DNA synthesis at oriC. An alternative function is suggested.

摘要

以微型染色体为模板在体外确定了前导链和后随链DNA的起始位点。通过与单链探针杂交分析复制中间体的片段。发现逆时针方向的前导链合成起始于最小起始点内或其附近（位置248至-44）。互补的后随链合成在oriC左侧的几个位置开始。结果还表明，在双链oricC DNA（前复制体）上，随着dnaA引导起始蛋白的组装，前导链和后随链进行协同合成。此外，DNA合成可以在位于asnA基因内且顺时针方向的蛋白质n'识别序列处起始。在n'位点的起始依赖于蛋白质i的活性，而oriC区域的前导链和后随链起始不受蛋白质i的影响。我们的结果表明，phi X174型引发体不参与oriC处不连续DNA合成的起始。提出了一种替代功能。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8ba/1167339/35bc1b712f6f/emboj00175-0327-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8ba/1167339/22e8da921ff4/emboj00175-0327-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8ba/1167339/35bc1b712f6f/emboj00175-0327-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8ba/1167339/22e8da921ff4/emboj00175-0327-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8ba/1167339/35bc1b712f6f/emboj00175-0327-b.jpg

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1
Initiation of Escherichia coli minichromosome replication at oriC and at protein n' recognition sites. Two modes for initiating DNA synthesis in vitro.大肠杆菌微型染色体在oriC和蛋白质n'识别位点处的复制起始。体外启动DNA合成的两种模式。
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本文引用的文献

1
Functional analysis of minichromosome replication: bidirectional and unidirectional replication from the Escherichia coli replication origin, oriC.微型染色体复制的功能分析：来自大肠杆菌复制起点oriC的双向和单向复制
J Bacteriol. 1980 Aug;143(2):1049-53. doi: 10.1128/jb.143.2.1049-1053.1980.
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Replication origin of the Escherichia coli K-12 chromosome: the size and structure of the minimum DNA segment carrying the information for autonomous replication.大肠杆菌K-12染色体的复制起点：携带自主复制信息的最小DNA片段的大小和结构
Mol Gen Genet. 1980 Apr;178(1):9-20. doi: 10.1007/BF00267207.
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Efficient in vitro replication of double-stranded DNA templates by a purified T4 bacteriophage replication system.
Philos Trans R Soc Lond B Biol Sci. 2004 Jan 29;359(1441):71-7. doi: 10.1098/rstb.2003.1366.
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DNA replication meets genetic exchange: chromosomal damage and its repair by homologous recombination.DNA复制与基因交换：染色体损伤及其同源重组修复
Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8461-8. doi: 10.1073/pnas.151260698.
5
Historical overview: searching for replication help in all of the rec places.历史概述：在所有推荐的地方寻找复制帮助。
Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8173-80. doi: 10.1073/pnas.131004998.
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The structure of the initiation complex at the replication origin, oriC, of Escherichia coli.大肠杆菌复制起点oriC处起始复合物的结构。
Nucleic Acids Res. 1993 Nov 11;21(22):5025-33. doi: 10.1093/nar/21.22.5025.
7
Escherichia coli PriA protein is essential for inducible and constitutive stable DNA replication.大肠杆菌PriA蛋白对于诱导型和组成型稳定DNA复制至关重要。
EMBO J. 1994 Nov 15;13(22):5338-45. doi: 10.1002/j.1460-2075.1994.tb06868.x.
8
Start sites for bidirectional in vitro DNA replication inside the replication origin, oriC, of Escherichia coli.大肠杆菌复制起点oriC内双向体外DNA复制的起始位点。
EMBO J. 1987 Aug;6(8):2469-72. doi: 10.1002/j.1460-2075.1987.tb02527.x.
9
Initiation of DNA replication in Escherichia coli.大肠杆菌中DNA复制的起始
J Bacteriol. 1987 Aug;169(8):3395-9. doi: 10.1128/jb.169.8.3395-3399.1987.
10
Transcripts within the replication origin, oriC, of Escherichia coli.大肠杆菌复制起点oriC内的转录本。
Nucleic Acids Res. 1987 Mar 25;15(6):2479-97. doi: 10.1093/nar/15.6.2479.
利用纯化的T4噬菌体复制系统对双链DNA模板进行高效体外复制。
J Biol Chem. 1980 May 10;255(9):4290-3.
4
Alkaline transfer of DNA to plastic membrane.DNA向塑料膜的碱性转移
Biochem Biophys Res Commun. 1984 Jul 18;122(1):340-4. doi: 10.1016/0006-291x(84)90480-7.
5
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6
The replication origin region of Escherichia coli: nucleotide sequence and functional units.大肠杆菌的复制起始区域：核苷酸序列与功能单元
Gene. 1983 Oct;24(2-3):265-79. doi: 10.1016/0378-1119(83)90087-2.
7
The 245 base-pair oriC sequence of the E. coli chromosome directs bidirectional replication at an adjacent region.大肠杆菌染色体的245个碱基对的oriC序列在相邻区域指导双向复制。
Nucleic Acids Res. 1983 May 11;11(9):2617-26. doi: 10.1093/nar/11.9.2617.
8
Chromosomal replication origin from the marine bacterium Vibrio harveyi functions in Escherichia coli: oriC consensus sequence.来自海洋细菌哈维氏弧菌的染色体复制起点在大肠杆菌中发挥作用：oriC共有序列。
Proc Natl Acad Sci U S A. 1983 Mar;80(5):1164-8. doi: 10.1073/pnas.80.5.1164.
9
Initiation signals for complementary strand DNA synthesis in the region of the replication origin of the Escherichia coli chromosome.大肠杆菌染色体复制起点区域中互补链DNA合成的起始信号。
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10
A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments.一对用于选择双酶切限制片段任一条DNA链的新型M13载体。
Gene. 1982 Oct;19(3):269-76. doi: 10.1016/0378-1119(82)90016-6.