Frequency of close positioning of chromosomal loci detected by FRET correlates with their participation in carcinogenic rearrangements in human cells.
Frequency of close positioning of chromosomal loci detected by FRET correlates with their participation in carcinogenic rearrangements in human cells.
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DOI：
10.1002/gcc.21988
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发表时间：
2012-11
影响因子：
3.7
通讯作者：
Nikiforov, Yuri E.
中科院分区：
文献类型：
--
作者：
Gandhi, Manoj;Evdokimova, Viktoria;Nikiforov, Yuri E.
已经证实，参与致癌基因重排的基因在人类细胞核中是非随机定位的，并且经常相互接近。然而，这些聚变伙伴之间的实际距离一直没有确定。荧光共振能量转移(FRET)现象是当供体荧光团靠近(~lt；10 nm)将部分能量转移到受体荧光团时观察到的。这项研究的目的是验证FRET在直接标记的DNA分子上的使用，以评估已知参与重排的基因之间在10 nm距离处定位的频率，并将其与它们经历重排的可能性相关联。在验证实验中，发现紧邻染色体区域的探针之间的FRET-SE(敏化发射)频率为93-96%，而位于大的线性分离上的随机基因座的探针之间的FRET-SE(敏化发射)的频率为0.1-0.2%。此外，我们还发现在甲状腺癌中形成重排的四对基因之间的FRET-SE频率在RET和CCDC6中为5%，在RET和NCOA4中为4%，在BRAF和AKAP9中为2%，在NTRK1和TPR中为2%。此外，FRET的出现频率与甲状腺癌中相应重排的发生率有很强的相关性(r=0.9871)。我们的发现表明，FRET可以作为一种技术来分析特定DNA区域之间的接近程度，并且在允许FRET的距离处基因定位的频率与它们经历染色体重排的可能性相关。
It has been well established that genes participating in oncogenic rearrangements are non-randomly positioned and frequently close to each other in human cell nuclei. However the actual distance between these fusion partners has never been determined. The phenomenon of Fluorescence Resonance Energy Transfer (FRET) is observed when a donor fluorophore is close (<10 nm) to transfer some of it energy to an acceptor fluorophore. The aim of this study was to validate the use of FRET on directly labeled DNA molecules to assess the frequency of positioning at <10 nm distances between genes known to be involved in rearrangement and to correlate it with their probability to undergo rearrangement. In the validation experiments, the frequency of FRET-SE (sensitized emission) was found to be 93-96% between probes for the immediately adjacent chromosomal regions as compared to 0.1-0.2% between probes for the random loci located on large linear separation. Further, we found that the frequency of FRET-SE between four pairs of genes that form rearrangements in thyroid cancer was 5% for RET and CCDC6, 4% for RET and NCOA4, 2% for BRAF and AKAP9, and 2% for NTRK1 and TPR. Moreover, the frequency with which FRET was observed showed strong correlation (r = 0.9871) with the prevalence of respective rearrangements in thyroid cancer. Our findings demonstrate that FRET can be used as a technique to analyze proximity between specific DNA regions and that the frequency of gene positioning at distances allowing FRET correlates with their probability to undergo chromosomal rearrangements.
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DOI：
10.1126/science.1178124
发表时间：
2009-11-27
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Mani RS;Tomlins SA;Callahan K;Ghosh A;Nyati MK;Varambally S;Palanisamy N;Chinnaiyan AM
通讯作者：
Chinnaiyan AM
DOI：
10.1073/pnas.0611503104
发表时间：
2007-02-20
影响因子：
11.1
作者：
Mukhopadhyay, Samrat;Krishnan, Rajaraman;Deniz, Ashok A.
通讯作者：
Deniz, Ashok A.
影响因子：
4.5
作者：
Lee, Ju-Han;Lee, Eung Seok;Chae, Yang-Seok
通讯作者：
Chae, Yang-Seok
影响因子：
3.4
作者：
Jakob, B.;Splinter, J.;Taucher-Scholz, G.
通讯作者：
Taucher-Scholz, G.
影响因子：
3.4
作者：
Gordon, GW;Berry, G;Herman, B
通讯作者：
Herman, B