Proteomics

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Crosslinking MS to elucidate the Architecture of the Nucleosome Remodelling and Deacetylase (NuRD) complex

ABSTRACT: The Nucleosome Remodeling and Deacetylase (NuRD) complex is essential for development in complex animals but has been refractory to detailed analysis because of its low abundance and resistance to recombinant production. In combination with other techniques, crosslinking mass spectrometry was used to elucidate the structure of the Nucleosome Remodelling and Deacetylase (NuRD) complex. Natively-purified NuRD, and four recombinantly-expressed NuRD subcomplexes, and a PWWP2A-MTA-HDAC-RBBP alternative ‘NuRD-like’ complex were subjected to crosslinking with DSS, ADH, BS3 and DMTMM.

INSTRUMENT(S): Q Exactive Plus, Q Exactive

ORGANISM(S): Homo Sapiens (human) Mus Musculus (mouse)

SUBMITTER: Jason Low  

LAB HEAD: Joel Mackay

PROVIDER: PXD010111 | Pride | 2021-01-08

REPOSITORIES: Pride

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150219_QEplus_BP_Jason_NuRD_Xlinkv6_F14.raw Raw
150219_QEplus_BP_Jason_NuRD_Xlinkv6_F15.raw Raw
150219_QEplus_BP_Jason_NuRD_Xlinkv6_pool.raw Raw
150219_QEplus_BP_Jason_NuRD_Xlinkv7_F14.raw Raw
150219_QEplus_BP_Jason_NuRD_Xlinkv7_F15_reinject.raw Raw
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Publications

The nucleosome remodeling and deacetylase (NuRD) complex is essential for metazoan development but has been refractory to biochemical analysis. We present an integrated analysis of the native mammalian NuRD complex, combining quantitative mass spectrometry, cross-linking, protein biochemistry, and electron microscopy to define the architecture of the complex. NuRD is built from a 2:2:4 (MTA, HDAC, and RBBP) deacetylase module and a 1:1:1 (MBD, GATAD2, and Chromodomain-Helicase-DNA-binding [CHD])  ...[more]

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