组蛋白H1对DNA和染色质体外复制的影响。

Influence of histone H1 on the in vitro replication of DNA and chromatin.

作者信息

Halmer L, Gruss C

机构信息

Fakultät für Biologie, Universität Konstanz, Germany.

出版信息

Nucleic Acids Res. 1995 Mar 11;23(5):773-8. doi: 10.1093/nar/23.5.773.

DOI:10.1093/nar/23.5.773

PMID:7708492

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC306758/

Abstract

The influence of histone H1 on DNA replication was studied using the SV40 in vitro replication system with two different templates: histone H1/DNA complexes and SV40 minichromosomes reconstituted with H1. We found that the cytosolic extracts used as a source of enzymes for in vitro replication contained high amounts of RNA which competed with template DNA for the binding of histone H1. Removal of this RNA was necessary to ensure the stability of the templates thus allowing for the first time the study of the replication of histone H1-carrying templates in vitro. In contrast to the inhibitory effect of histone H1 on the initiation of transcription, bound H1, when present at physiological ratios, does not interfere with the in vitro replication of DNA and minichromosomes. Ratios higher than one H1 molecule per nucleosome affected replication of reconstituted SV40 minichromosomes.

摘要

利用SV40体外复制系统，以两种不同模板研究了组蛋白H1对DNA复制的影响：组蛋白H1/DNA复合物和用H1重建的SV40微型染色体。我们发现，用作体外复制酶源的胞质提取物含有大量RNA，这些RNA与模板DNA竞争组蛋白H1的结合。去除这种RNA对于确保模板的稳定性是必要的，从而首次能够在体外研究携带组蛋白H1的模板的复制。与组蛋白H1对转录起始的抑制作用相反，当以生理比例存在时，结合的H1不会干扰DNA和微型染色体的体外复制。高于每个核小体一个H1分子的比例会影响重建的SV40微型染色体的复制。