DNA Binding Restricts the Intrinsic Conformational Flexibility of Methyl CpG Binding Protein 2 (MeCP2)

Hansen, Jeffrey C.; Wexler, Brian B.; Rogers, Danielle J.; Hite, Kristopher C.; Panchenko, Tanya; Ajith, Sandya; Black, Ben E.

doi:10.1074/jbc.m111.234609

Cited by 28 publications

(32 citation statements)

References 54 publications

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“…Instead, we observe that that both the helical histone fold and unstructured tail regions exchange rapidly, with all of H2A and most of H2B reaching 75–100% D in only 10 seconds (Figure 2, left). This rapid rate of exchange is similar to that observed for intrinsically disordered proteins such as MeCP2 (Hansen et al, 2011). Intrinsically disordered proteins contain few H-bonds as they lack stable secondary and tertiary structure.…”

Section: Resultssupporting

confidence: 81%

Chaperone Nap1 Shields Histone Surfaces Used in a Nucleosome and Can Put H2A-H2B in an Unconventional Tetrameric Form

et al. 2013

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Summary The histone H2A-H2B heterodimer is an integral component of the nucleosome. The cellular localization and deposition of H2A-H2B into chromatin is regulated by numerous factors including histone chaperones such as Nucleosome Assembly Protein 1 (Nap1). We use hydrogen-deuterium exchange coupled to mass spectrometry to characterize H2A-H2B and Nap1. Unexpectedly, we find that at low ionic strength the α-helices in H2A-H2B are frequently sampling partially disordered conformations, and that binding to Nap1 reduces this conformational sampling. We identify the interaction surface between H2A-H2B and Nap1, and confirm its relevance both in vitro and in vivo. We show that two copies of H2A-H2B bound to a Nap1 homodimer form a tetramer with contacts between H2B chains similar to those in the four-helix bundle structural motif. The organization of the complex reveals that Nap1 competes with histone-DNA and inter-histone interactions observed in the nucleosome, thereby regulating the availability of histones for chromatin assembly.

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Section: Resultssupporting

confidence: 81%

Chaperone Nap1 Shields Histone Surfaces Used in a Nucleosome and Can Put H2A-H2B in an Unconventional Tetrameric Form

et al. 2013

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“…Hydrogen-deuterium exchange (H/DX) is a powerful solution-based approach to access information about protein structure, dynamics, and folding (Englander, 2006). We and others have coupled H/DX to mass spectrometry (MS) in order to access dynamic information on macromolecular interactions, providing key insight to complement known static structures (Hansen et al, 2011; Mendillo et al, 2009; Lee et al, 2004; Panchenko et al, 2011). In order to determine how HJURP binding affects CENP-A/H4 dynamics, we measured levels of CENP-A/H4 protection in complex with HJURP relative to independent measurements on (CENP-A/H4) 2 heterotetramers using the conditions optimized for CENP-A/H4 proteolysis and peptide recovery at the experimental steps following H/DX (Black et al, 2004).…”

Section: Resultsmentioning

confidence: 99%

HJURP Uses Distinct CENP-A Surfaces to Recognize and to Stabilize CENP-A/Histone H4 for Centromere Assembly

et al. 2012

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Summary Centromeres are defined by the presence of chromatin containing the histone H3 variant, CENP-A, whose assembly into nucleosomes requires the chromatin assembly factor HJURP. We find that while surface-exposed residues in the CENP-A targeting domain (CATD) are the primary sequence determinants for HJURP recognition, buried CATD residues that generate rigidity with H4 are also required for efficient incorporation into centromeres. HJURP contact points adjacent to the CATD on the CENP-A surface are not used for binding specificity but rather to transmit stability broadly throughout the histone fold domains of both CENP-A and H4. Further, an intact CENP-A/CENP-A interface is a requirement for stable chromatin incorporation immediately upon HJURP-mediated assembly. These data offer insight into the mechanism by which HJURP discriminates CENP-A from bulk histone complexes and chaperones CENP-A/H4 for a substantial portion of the cell cycle prior to mediating chromatin assembly at the centromere.

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“…Folded regions (e.g., α-helices and β-sheets) only exchange upon transient unfolding events when amide protons lose main chain hydrogen bonding. Slow exchange can be achieved by many stabilizing interactions (23), including, in the case of DNA-binding proteins, assembly into higher-order complexes with DNA (4,24,25). In order to test the extent to which the polypeptide backbone dynamics of the structured histone core of the nucleosomal subunits are altered by array folding, we monitored H/DX exchange behavior of folded and unfolded array fibers (time points at 10, 100, 1,000, and 10,000 s at 23°C) (Fig.…”

Section: Resultsmentioning

confidence: 99%

Replacement of histone H3 with CENP-A directs global nucleosome array condensation and loosening of nucleosome superhelical termini

Panchenko

Sorensen

Woodcock

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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Centromere protein A (CENP-A) is a histone H3 variant that marks centromere location on the chromosome. To study the subunit structure and folding of human CENP-A-containing chromatin, we generated a set of nucleosomal arrays with canonical core histones and another set with CENP-A substituted for H3. At the level of quaternary structure and assembly, we find that CENP-A arrays are composed of octameric nucleosomes that assemble in a stepwise mechanism, recapitulating conventional array assembly with canonical histones. At intermediate structural resolution, we find that CENP-A-containing arrays are globally condensed relative to arrays with the canonical histones. At high structural resolution, using hydrogen-deuterium exchange coupled to mass spectrometry (H/DX-MS), we find that the DNA superhelical termini within each nucleosome are loosely connected to CENP-A, and we identify the key amino acid substitution that is largely responsible for this behavior. Also the C terminus of histone H2A undergoes rapid hydrogen exchange relative to canonical arrays and does so in a manner that is independent of nucleosomal array folding. These findings have implications for understanding CENP-A-containing nucleosome structure and higher-order chromatin folding at the centromere.hydrogen exchange | epigenetics

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DNA Binding Restricts the Intrinsic Conformational Flexibility of Methyl CpG Binding Protein 2 (MeCP2)

Cited by 28 publications

References 54 publications

Chaperone Nap1 Shields Histone Surfaces Used in a Nucleosome and Can Put H2A-H2B in an Unconventional Tetrameric Form

Chaperone Nap1 Shields Histone Surfaces Used in a Nucleosome and Can Put H2A-H2B in an Unconventional Tetrameric Form

HJURP Uses Distinct CENP-A Surfaces to Recognize and to Stabilize CENP-A/Histone H4 for Centromere Assembly

Replacement of histone H3 with CENP-A directs global nucleosome array condensation and loosening of nucleosome superhelical termini

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