Iarovaia Olga V, Shkumatov Petr, Razin Sergey V
Laboratory of Structural and Functional Organization of Chromosomes, Institute of Gene Biology RAS, Vavilov Street 34/5, 119334, Moscow, Russia.
J Cell Sci. 2004 Sep 1;117(Pt 19):4583-90. doi: 10.1242/jcs.01332.
The spatial organization in interphase nuclei of the breakpoint cluster regions (BCRs) of the AML-1 and ETO genes frequently participating in reciprocal t(8;21) translocations was studied using cytological and biochemical approaches. Both BCRs were found to be localized preferentially, but not exclusively, to the nuclear matrix, as shown by hybridization of specific probes with nuclear halos. This association was not related to transcription, because the transcribed regions of both genes located far from BCRs were located preferentially in loop DNA, as shown by in situ hybridization. The sites of association with the nuclear matrix of the intensely transcribed AML-1 gene were mapped also using the biochemical PCR-based approach. Only the BCR was found to be associated with the nuclear matrix, whereas the other transcribed regions of this gene turned out to be positioned randomly in respect to the nuclear matrix. The data are discussed in the framework of the hypothesis postulating that the nuclear matrix plays an important role in determining the positions of recombination-prone areas.
运用细胞学和生化方法,研究了急性髓性白血病1(AML-1)基因和八分体-21(ETO)基因的断裂簇区域(BCR)在间期细胞核中的空间组织,这两个基因常参与相互易位t(8;21)。通过特异性探针与核晕的杂交显示,两个BCR均优先定位于核基质,但并非唯一。这种关联与转录无关,因为原位杂交显示,这两个基因远离BCR的转录区域优先位于环状DNA中。还使用基于生化聚合酶链反应(PCR)的方法绘制了高转录AML-1基因与核基质的结合位点。结果发现,只有BCR与核基质相关,而该基因的其他转录区域相对于核基质的定位是随机的。在假设核基质在确定易重组区域位置中起重要作用的框架内,对这些数据进行了讨论。