García-Vilchis David, Aranda-Anzaldo Armando
Facultad de Medicina, Laboratorio de Biología Molecular y Neurociencias, Universidad Autónoma del Estado de México, Toluca 50180, Edo. Méx., Mexico.
J Cell Biochem. 2017 Dec;118(12):4487-4497. doi: 10.1002/jcb.26106. Epub 2017 May 31.
Classical observations have shown that during the interphase the chromosomal DNA of metazoans is organized in supercoiled loops attached to a compartment known as the nuclear matrix (NM). Fragments of chromosomal DNA able to bind the isolated NM in vitro are known as matrix associated/attachment/addressed regions or MARs. No specific consensus sequence or motif has been found that may constitute a universal, defining feature of MARs. On the other hand, high-salt resistant DNA-NM interactions in situ define true DNA loop anchorage regions or LARs, that might correspond to a subset of the potential MARs but are not necessarily identical to MARs characterized in vitro, since there are several examples of MARs able to bind the NM in vitro but which are not actually bound to the NM in situ. In the present work we assayed the capacity of two LARs, as well as of shorter fragments within such LARs, for binding to the NM in vitro. Paradoxically the isolated (≈2 kb) LARs cannot bind to the NM in vitro while their shorter (≈300 pb) sub-fragments and other non-related but equally short DNA fragments, bind to the NM in a high-salt resistant fashion. Our results suggest that the ability of a given DNA fragment for binding to the NM in vitro primarily depends on the length of the fragment, suggesting that binding to the NM is modulated by the local topology of the DNA fragment in suspension that it is known to depend on the DNA length. J. Cell. Biochem. 118: 4487-4497, 2017. © 2017 Wiley Periodicals, Inc.
经典观察表明,在间期,后生动物的染色体DNA以超螺旋环的形式组织,附着于一个称为核基质(NM)的隔室。能够在体外与分离的核基质结合的染色体DNA片段被称为基质相关/附着/靶向区域或MARs。尚未发现可能构成MARs普遍定义特征的特定共有序列或基序。另一方面,原位耐高盐的DNA-核基质相互作用定义了真正的DNA环锚定区域或LARs,其可能对应于潜在MARs的一个子集,但不一定与体外鉴定的MARs相同,因为有几个MARs的例子,它们能够在体外与核基质结合,但实际上在原位并未与核基质结合。在本研究中,我们检测了两个LARs以及这些LARs内较短片段在体外与核基质结合的能力。矛盾的是,分离的(≈2 kb)LARs在体外不能与核基质结合,而它们较短的(≈300 pb)亚片段和其他不相关但同样短的DNA片段,则以耐高盐的方式与核基质结合。我们的结果表明,给定DNA片段在体外与核基质结合的能力主要取决于片段的长度,这表明与核基质的结合受悬浮液中DNA片段的局部拓扑结构调节,而局部拓扑结构已知取决于DNA长度。《细胞生物化学杂志》118: 4487 - 4497, 2017年。© 2017威利期刊公司