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Role of the M-loop and reactive center loop domains in the folding and bridging of nucleosome arrays by MENT

By Evelyn M. Springhetti, Natalia E. Istomina, James C. Whisstock, Tatiana Nikitina, Chris L. Woodcock and Sergei A. Grigoryev

Abstract

MENT is a developmentally regulated heterochromatin-associated protein that condenses chromatin in terminally differentiated avian blood cells. Its homology to the serpin protein family suggests that the conserved serpin reactive center loop (RCL) and the unique M-loop are important for its function. To examine the role of these domains, we studied the interaction of wild-type and mutant MENT with naked DNA and biochemically defined nucleosome arrays reconstituted from 12-mer repeats containing nucleosome positioning sequences. Wild-type MENT folded the naked DNA duplexes into closely juxtaposed parallel structures (“tramlines”). Deletion of the M-loop, but not inactivation of the RCL, prevented tramline formation and the cooperative interaction of MENT with DNA. Reconstitution of wildtyp

Year: 2003
OAI identifier: oai:CiteSeerX.psu:10.1.1.134.622
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