replication independent formation of extrachromosomal circular dna in mammalian cell-free system染色体外环状dna的复制独立形成颗粒系统的哺乳动物.pdfVIP

Replication Independent Formation of Extrachromosomal Circular DNA in Mammalian Cell-Free System Zoya Cohen, Sara Lavi* Department of Cell Research and Immunology, Tel Aviv University, Tel Aviv, Israel Abstract Extrachromosomal circular DNA (eccDNA) is a pool of circular double stranded DNA molecules found in all eukaryotic cells and composed of repeated chromosomal sequences. It was proposed to be involved in genomic instability, aging and alternative telomere lengthening. Our study presents novel mammalian cell-free system for eccDNA generation. Using purified protein extract we show that eccDNA formation does not involve de-novo DNA synthesis suggesting that eccDNA is generated through excision of chromosomal sequences. This process is carried out by sequence- independent enzymes as human protein extract can produce mouse- specific eccDNA from high molecular weight mouse DNA, and vice versa. EccDNA production does not depend on ATP, requires residual amounts of Mg2+ and is enhanced by double strand DNA breaks. Citation: Cohen Z, Lavi S (2009) Replication Independent Formation of Extrachromosomal Circular DNA in Mammalian Cell-Free System. PLoS ONE 4(7): e6126. doi:10.1371/journal.pone.0006126 Editor: Beth A. Sullivan, Duke University, United States of America Received March 19, 2009; Accepted May 29, 2009; Published July 1, 2009 Copyright: 2009 Cohen et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This research was funded by the Israel Cancer Association through the estate of the late Ethel Cohen. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of