|
|
|
| Comparison of ARS Element Forming Nucleosome on Saccharomyces cerevisiae YPH499 Ⅲ Chromosome in vitro |
| ZHAO Xiu-juan, ZHENG Xiao-dong, XUE Tao-tao, CAI Lu |
| School of Mathematics Physics and Biological Engineering, Inner Mongolia University of Science and Technology, Baotou 014010, China |
|
|
|
Abstract The regulation of DNA replication initiation sites has a guarantee for DNA replication and cell division. Nucleosome positioning is an important factor for the regulation of DNA replication initiation sites. But it is not clear that how nucleosome positioning regulates DNA replication initiation. It has important biological significance to understand the mechanism of DNA replication initiation. In the study, ten autonomously replicating sequences(ARS) of Saccharomyces cerevisiae YPH499 Ⅲ chromosome were divided into efficient and inactivity groups. These aim sequences were amplificated by PCR, then recycled and purified using TIANgel Maxi Purification kit. A gradient salt dialysis method was employed to assemble nucleosome in vitro, then nucleosome depended on the purpose sequence were analyzed with Biotin labeling method. The ability of forming nucleosome was analyzed with Image J software.The result showed that nucleosome assembly strength which was analysised by Image J software is respectively 0.706, 1.395, 2.989, 0, 0.031, 0, 0, 0.714, 0 and 0 from ARS302 to ARS315. In other words, nucleosome assembly strength was as follows:ARS304>ARS303>ARS313>ARS302>ARS306>ARS314,ARS305,ARS307,ARS309,ARS315. The results indicated the inactive ARS were more likely to form nucleosome than the efficient ARS; Replication initiation sites often show anucleosome-depleted region.
|
|
Received: 20 September 2014
Published: 25 November 2014
|
|
|
|
|
|
[1] Lee W, Tillo D, Bray N, et al. A high-resolution atlas of nucleosome occupancy in yeast. Nat Genet, 2007,39(10):1235-1244.
[2] Audit B, Zaghloul L, Vaillant C, et al. Open chromatin encoded in DNA sequence is the signature of 'master' replication origins in human cells. Nucleic Acids Res, 2009,37(18):6064-6075.
[3] Hagerman K A, Ruan H, Edamura K N, et al. The ATTCT repeats of spinocerebellar ataxia type 10 display strong nucleosome assembly which is enhanced by repeat interruptions. Gene, 2009,434(1-2):29-34.
[4] Jiang C, Pugh B F. A compiled and systematic reference map of nucleosome positions across the Saccharomyces cerevisiae genome. Genome Biol, 2009,10(10):R109.
[5] Theis J F, Dershowitz A, Irene C, et al. Identification of mutations that decrease the stability of a fragment of Saccharomyces cerevisiae chromosome III lacking efficient replicators. Genetics, 2007,177(3):1445-1458.
[6] Irene C, Maciariello C, Cioci F, et al. Identification of the sequences required for chromosomal replicator function in Kluyveromyces lactis. Mol Microbiol, 2004,51(5):1413-1423.
[7] Theis J F, Newlon C S. Two compound replication origins in Saccharomyces cerevisiae contain redundant origin recognition complex binding sites. Mol Cell Biol, 2001,21(8):2790-2801.
[8] Simpson R T. Nucleosome positioning can affect the function of a cis-acting DNA element in vivo. Nature, 1990,343(6256):387-389.
[9] Lipford J R, Bell S P. Nucleosomes positioned by ORC facilitate the initiation of DNA replication. Mol Cell, 2001,7(1):21-30.
[10] Eaton M L, Galani K, Kang S, et al. Conserved nucleosome positioning defines replication origins. Genes Dev, 2010,24(8):748-753.
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
