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

doi:10.1083/jcb.137.2.263

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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 complexis involved in the remodeling of chromatin during gene activation.We have used antibodies specific against three human subunitsof this complex to study its subnuclear localization, as wellas its potential association with active chromatin and the nuclear skeleton. Immunofluorescence studies revealed a punctatenuclear labeling pattern that was excluded from the nucleoliand from regions of condensed chromatin. Dual labeling failedto reveal significant colocalization of BRG1 or hBRM proteinswith RNA polymerase II or with nuclear speckles involved insplicing. Chromatin fractionation experiments showed that both soluble and insoluble active chromatin are enriched in thehSWI/SNF proteins as compared with bulk chromatin. hSWI/SNFproteins were also found to be associated with the nuclear matrixor nuclear scaffold, suggesting that a fraction of the hSWI/SNFcomplex 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 postdoctoralfellowships. The work was supported by Association pour la Recherchesur le Cancer, Ligue Nationale Française Contre le Cancer,and the Actions Concertées des Sciences du Vivant programof the French Ministry of Science.

Please address all correspondence to Moshe Yaniv, Unitédes 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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