Smith H C, Berezney R, Brewster J M, Rekosh D
Biochemistry. 1985 Feb 26;24(5):1197-202. doi: 10.1021/bi00326a022.
The association of adenoviral DNA with the high salt (2 M NaCl) resistant nuclear fraction, termed the nuclear matrix, has been investigated in HeLa cells at different times after infection with adenovirus type 5. When nuclear matrices were prepared in the absence of exogenously added nucleases, Ad5 DNA was quantitively associated with the matrix throughout the infection period (0-24 h). Moreover, early in infection (0-10 h) Ad5 DNA was severalfold enriched in DNase I digested nuclear matrices (8-15% of total nuclear DNA) compared to the high salt soluble chromatin fraction (85-90% of total nuclear DNA). At later times after infection, progressively more Ad5 DNA appeared in the chromatin fraction until, at 24 h, the nuclear matrix was strikingly depleted in Ad5 DNA. A large proportion of the Ad5 DNA in nuclear matrices prepared early in infection, e.g., 4 h, was full length in size. At later times (12-24 h) most of the viral DNA was fragmented to a size equivalent to total matrix DNA (100-1000 base pairs). The apparent switch of the matrix-associated viral DNA from a relatively DNase I resistant to sensitive state was initiated approximately at the time when viral DNA replication began (12 h). Since no discrete portion of the Ad5 genome was significantly enriched at the sites of attachment to the nuclear matrix throughout the infection period, the switch in DNase I sensitivity is not mediated by a change in DNA sequence attachment to the matrix.(ABSTRACT TRUNCATED AT 250 WORDS)
在感染5型腺病毒后的不同时间,对HeLa细胞中腺病毒DNA与称为核基质的高盐(2M NaCl)抗性核组分之间的关联进行了研究。当在不添加外源核酸酶的情况下制备核基质时,在整个感染期(0 - 24小时)Ad5 DNA与基质定量相关。此外,在感染早期(0 - 10小时),与高盐可溶性染色质组分(占总核DNA的85 - 90%)相比,Ad5 DNA在经DNase I消化的核基质中富集了几倍(占总核DNA的8 - 15%)。在感染后的后期,越来越多的Ad5 DNA出现在染色质组分中,直到24小时时,核基质中的Ad5 DNA明显减少。在感染早期(例如4小时)制备的核基质中,很大一部分Ad5 DNA大小为全长。在后期(12 - 24小时),大多数病毒DNA被片段化至与总基质DNA相当的大小(100 - 1000个碱基对)。与基质相关的病毒DNA从相对抗DNase I状态到敏感状态的明显转变大约在病毒DNA复制开始时(12小时)启动。由于在整个感染期,Ad5基因组的任何离散部分在与核基质附着的位点都没有显著富集,DNase I敏感性的转变不是由与基质附着的DNA序列变化介导的。(摘要截短于250字)