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© The Rockefeller University Press, 0021-9525/1997//263 $5.00
The Journal of Cell Biology, Volume 137, Number 2, , 1997 263-274

Article

Components of the Human SWI/SNF Complex Are Enriched in Active Chromatin and Are Associated with the Nuclear Matrix



Jose C. Reyes, Christian Muchardt, and Moshe Yaniv

Unité des Virus Oncogènes, UA1644 du Centre National de la Recherche Scientifique, Département des Biotechnologies, Institut Pasteur, 75724 Paris, France

Biochemical and genetic evidence suggest that the SWI/SNF complex is involved in the remodeling of chromatin during gene activation. We have used antibodies specific against three human subunits of this complex to study its subnuclear localization, as well as its potential association with active chromatin and the nuclear skeleton. Immunofluorescence studies revealed a punctate nuclear labeling pattern that was excluded from the nucleoli and from regions of condensed chromatin. Dual labeling failed to reveal significant colocalization of BRG1 or hBRM proteins with RNA polymerase II or with nuclear speckles involved in splicing. Chromatin fractionation experiments showed that both soluble and insoluble active chromatin are enriched in the hSWI/SNF proteins as compared with bulk chromatin. hSWI/SNF proteins were also found to be associated with the nuclear matrix or nuclear scaffold, suggesting that a fraction of the hSWI/SNF complex could be involved in the chromatin organization properties associated with matrix attachment regions.

Abbreviations used in this paper: CTD, carboxy-terminal domain; DAPI, 4',6-diamidino-2-phenylindole; GR, glucocorticoid receptor; LIS, 3,5-diiodosalicilic acid, lithium salt; MAR, matrix-associated region; SAR, scaffold-associated region; Rb, retinoblastoma; RNAP II, RNA polymerase II.

J.C. Reyes was the recipient of European Molecular Biology Organization and Human Frontier Science Program Organization postdoctoral fellowships. The work was supported by Association pour la Recherche sur le Cancer, Ligue Nationale Française Contre le Cancer, and the Actions Concertées des Sciences du Vivant program of the French Ministry of Science.

Please address all correspondence to Moshe Yaniv, Unité des Virus Oncogènes, Département des Biotechnologies, 25, rue Dr. Roux, 75724 Paris Cedex 15, France. Tel.: (33) 1-45-68-85-13. Fax: (33) 1-40-61-30-33.


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