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一种细胞DNA复制蛋白与腺病毒DNA复制起点的特异性结合。

Specific binding of a cellular DNA replication protein to the origin of replication of adenovirus DNA.

作者信息

Nagata K, Guggenheimer R A, Hurwitz J

出版信息

Proc Natl Acad Sci U S A. 1983 Oct;80(20):6177-81. doi: 10.1073/pnas.80.20.6177.

DOI:10.1073/pnas.80.20.6177
PMID:6336326
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC394258/
Abstract

Nuclear factor I, a 47-kilodalton protein, purified from nuclear extracts of uninfected HeLa cells, is involved in the initiation and possibly the elongation of replicating adenovirus (Ad) DNA in vitro. The binding of nuclear factor I to DNA has been monitored by a filter binding assay of nuclear factor I to DNA has been monitored by a filter binding assay using plasmid pLA1 DNA, which contains a 3,290 base-pair fragment derived from the left-hand terminus (coordinates, 0-9.4 map units) of Ad serotype 5 DNA. Nuclear factor I binds selectively to a double-stranded fragment spanning nucleotides 0-451 to the Ad genome. The retention of the 451-base-pair DNA fragment-nuclear factor I complex on nitrocellulose filters does not require Mg2+ or ATP and is resistant to high ionic strength. DNase I protection experiments revealed that nuclear factor I binds to a nucleotide sequence located at position 17-48, close to the terminus of Ad DNA. This 32-nucleotide sequence contains four "consensus" sequences present in various serotypes of Ad DNA and is capable of forming higher ordered structures. The role of nuclear factor I and this DNA sequence in the generation of Ad preterminal protein-dCMP initiation complex is discussed.

摘要

核因子I是一种47千道尔顿的蛋白质，从未感染的HeLa细胞核提取物中纯化得到，它参与体外复制腺病毒（Ad）DNA的起始过程，可能还参与延伸过程。通过使用质粒pLA1 DNA的滤膜结合试验监测核因子I与DNA的结合，该质粒包含一个3290碱基对的片段，源自腺病毒5型DNA左手末端（坐标为0 - 9.4图谱单位）。核因子I选择性地结合到腺病毒基因组中跨越核苷酸0 - 451的双链片段上。451碱基对的DNA片段 - 核因子I复合物在硝酸纤维素滤膜上的保留不需要Mg2+或ATP，并且对高离子强度具有抗性。DNase I保护实验表明，核因子I结合到位于腺病毒DNA末端附近位置17 - 48的核苷酸序列上。这个32核苷酸序列包含存在于各种腺病毒DNA血清型中的四个“共有”序列，并且能够形成更高阶的结构。本文讨论了核因子I和这个DNA序列在腺病毒末端前蛋白 - dCMP起始复合物形成中的作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e62/394258/10533f735e0a/pnas00646-0040-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e62/394258/638830e4d28e/pnas00646-0038-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e62/394258/6ec6120f63a3/pnas00646-0038-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e62/394258/8cb1b3a66392/pnas00646-0039-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e62/394258/58cd30ffed0e/pnas00646-0039-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e62/394258/10533f735e0a/pnas00646-0040-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e62/394258/638830e4d28e/pnas00646-0038-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e62/394258/6ec6120f63a3/pnas00646-0038-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e62/394258/8cb1b3a66392/pnas00646-0039-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e62/394258/58cd30ffed0e/pnas00646-0039-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e62/394258/10533f735e0a/pnas00646-0040-a.jpg

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Proc Natl Acad Sci U S A. 1982 Apr;79(7):2221-5. doi: 10.1073/pnas.79.7.2221.
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Fluorescent protein tagging of adenoviral proteins pV and pIX reveals 'late virion accumulation compartment'.荧光蛋白标记腺病毒蛋白 pV 和 pIX 揭示“晚期病毒粒子积累隔室”。
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Atomic-resolution mapping of transcription factor-DNA interactions by femtosecond laser crosslinking and mass spectrometry.飞秒激光交联和质谱法原子分辨率绘制转录因子-DNA 相互作用图谱。
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