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猿猴病毒40插入突变体的DNA复制与染色质结构

DNA replication and chromatin structure of simian virus 40 insertion mutants.

作者信息

Innis J W, Scott W A

出版信息

Mol Cell Biol. 1984 Aug;4(8):1499-507. doi: 10.1128/mcb.4.8.1499-1507.1984.

DOI:10.1128/mcb.4.8.1499-1507.1984
PMID:6092914
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC368940/
Abstract

Insertion of DNA segments into the nuclease-sensitive region of simian virus 40 alters both replication efficiency and chromatin structure. Mutants containing large insertions between the simian virus 40 origin of replication (ori site) and the 21-base-pair repeated sequences replicated poorly when assayed by transfection into COS-1 cells. Replication of mutants with shorter insertions was moderately reduced. This effect was cis-acting and independent of the nucleotide sequence of the insert. The nuclease-sensitive chromatin structure was retained in these mutants, but the pattern of cleavage sites was displaced in the late direction from the ori site. New cleavage sites appeared within the inserted sequences, suggesting that information specifying the nuclease-sensitive chromatin structure is located on the late side of the inserts. Accessibility to BglI (which cleaves within the ori site) was reduced in the larger insertion mutants. These results support the conclusion that efficient function of the viral origin of replication is correlated with its proximity to an altered chromatin structure.

摘要

将DNA片段插入猴病毒40的核酸酶敏感区域会改变复制效率和染色质结构。当通过转染到COS-1细胞中进行检测时，在猴病毒40复制起点（ori位点）和21个碱基对重复序列之间含有大的插入片段的突变体复制能力很差。插入片段较短的突变体的复制能力则适度降低。这种效应是顺式作用的，且与插入片段的核苷酸序列无关。这些突变体中保留了核酸酶敏感的染色质结构，但切割位点的模式从ori位点向晚期方向发生了位移。在插入序列内出现了新的切割位点，这表明指定核酸酶敏感染色质结构的信息位于插入片段的晚期一侧。在较大插入片段的突变体中，BglI（在ori位点内切割）的可及性降低。这些结果支持了这样的结论，即病毒复制起点的有效功能与其接近改变的染色质结构相关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8f2/368940/0690b97d4e54/molcellb00150-0085-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8f2/368940/bc06f3214601/molcellb00150-0082-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8f2/368940/98151cdd8974/molcellb00150-0082-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8f2/368940/b0960f3f92c1/molcellb00150-0083-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8f2/368940/cd13296faa45/molcellb00150-0083-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8f2/368940/f503c883594f/molcellb00150-0084-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8f2/368940/ebb1d8c8b489/molcellb00150-0084-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8f2/368940/0690b97d4e54/molcellb00150-0085-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8f2/368940/bc06f3214601/molcellb00150-0082-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8f2/368940/98151cdd8974/molcellb00150-0082-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8f2/368940/b0960f3f92c1/molcellb00150-0083-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8f2/368940/cd13296faa45/molcellb00150-0083-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8f2/368940/f503c883594f/molcellb00150-0084-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8f2/368940/ebb1d8c8b489/molcellb00150-0084-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8f2/368940/0690b97d4e54/molcellb00150-0085-a.jpg

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本文引用的文献

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Chromatin structure and gene activity: the role of nonhistone chromosomal proteins.染色质结构与基因活性：非组蛋白染色体蛋白的作用
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Induction of altered chromatin structures by simian virus 40 enhancer and promoter elements.猿猴病毒40增强子和启动子元件诱导染色质结构改变
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Chromatin structure of simian virus 40-pBR322 recombinant plasmids in COS-1 cells.猿猴病毒40-pBR322重组质粒在COS-1细胞中的染色质结构
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J Virol. 1987 Jul;61(7):2143-9. doi: 10.1128/JVI.61.7.2143-2149.1987.
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Functional analysis of the role of the A + T-rich region and upstream flanking sequences in simian virus 40 DNA replication.富含A+T区域及上游侧翼序列在猴病毒40 DNA复制中作用的功能分析。
Mol Cell Biol. 1986 Dec;6(12):4570-7. doi: 10.1128/mcb.6.12.4570-4577.1986.
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Bidirectional promoter elements of simian virus 40 are required for efficient replication of the viral DNA.猴病毒40的双向启动子元件是病毒DNA高效复制所必需的。
Mol Cell Biol. 1986 Oct;6(10):3513-22. doi: 10.1128/mcb.6.10.3513-3522.1986.
Mol Cell Biol. 1983 Dec;3(12):2203-10. doi: 10.1128/mcb.3.12.2203-2210.1983.
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The promoter-specific transcription factor Sp1 binds to upstream sequences in the SV40 early promoter.启动子特异性转录因子Sp1与SV40早期启动子中的上游序列结合。
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Simian virus 40 early- and late-region promoter functions are enhanced by the 72-base-pair repeat inserted at distant locations and inverted orientations.猿猴病毒40早期和晚期区域启动子功能可通过插入到远处位置且方向相反的72碱基对重复序列得到增强。
Mol Cell Biol. 1983 Jun;3(6):991-9. doi: 10.1128/mcb.3.6.991-999.1983.
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The repeated GC-rich motifs upstream from the TATA box are important elements of the SV40 early promoter.TATA框上游富含GC的重复基序是SV40早期启动子的重要元件。
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Definition of the simian virus 40 early promoter region and demonstration of a host range bias in the enhancement effect of the simian virus 40 72-base-pair repeat.猿猴病毒40早期启动子区域的定义以及猿猴病毒40 72碱基对重复序列增强效应中宿主范围偏向性的证明。
Proc Natl Acad Sci U S A. 1983 Feb;80(3):721-5. doi: 10.1073/pnas.80.3.721.
8
SV40 deletion mutants lacking the 21-bp repeated sequences are viable, but have noncomplementable deficiencies.缺乏21个碱基对重复序列的SV40缺失突变体是可行的，但存在不可互补的缺陷。
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High efficiency polyoma DNA transfection of chloroquine treated cells.氯喹处理细胞的高效多瘤病毒DNA转染
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Deletion mapping of DNA regions required for SV40 early region promoter function in vivo.体内SV40早期区域启动子功能所需DNA区域的缺失图谱分析。
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