Interaction of MAR‐sequences with nuclear matrix proteins

The recent discovery of DNA sequences responsible for the specific attachment of chromosomal DNA to the nuclear skeleton (MARs/SARs) was an important step towards our understanding of the functional and structural organization of eukaryotic chromatin [Mirkovitch et al.: Cell 44:273–282, 1984; Cockerill and Garrard: Cell 44:273–282, 1986]. A most important question, however, remains the nature of the matrix proteins involved in the specific binding of the MARs. It has been shown that topoisomerase II and histone H1 were capable of a specific interaction with SARs by the formation of precipitable complexes [Adachi et al.; EMBO J 8:3997–4006, 1989; lzaurralde et al.: J Mol Biol 210:573–585, 1989]. Here, applying a different approach, we were able to “visualize” some of the skeletal proteins recognizing and specifically binding MAR‐sequences. It is shown that the major matrix proteins are practically the same in both salt‐ and LIS‐extracted matrices. However, the relative MAR‐binding activity of the individual protein components may be different, depending on the method of matrix preparation. The immunological approach applied here allowed us to identify some of the individual MAR‐binding matrix proteins. Histone H1 and nuclear actin are shown to be not only important components of the matrix, but to be involved in a highly efficient interaction with MAR‐sequences as well. Evidence is presented that proteins recognized by the anti‐HMG antibodies also participate in Mar‐interactions. © 1992 Wiley‐Liss, Inc.