DNA Length Modulates the Affinity of Fragments of Genomic DNA for the Nuclear Matrix In Vitro

4 years ago

DNA Length Modulates the Affinity of Fragments of Genomic DNA for the Nuclear Matrix In Vitro

David García-Vilchis, Armando Aranda-Anzaldo

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. DNA loop attachment regions (LARs) ∼2 kb long, naturally bound in situ to the nuclear matrix (NM), cannot bind to the NM in vitro when isolated from their chromosomal context. In contrast, short (∼300 bp) but varied sub-fragments derived from such LARs or from non-related genomic DNA regions, bind to the NM in vitro. Our results suggest 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.

Publisher URL: http://onlinelibrary.wiley.com/resolve/doi

DOI: 10.1002/jcb.26106