Identification of novel Sin3A ‘proximate’ proteins and substrates using Biotin-ID (360G-Wellcome-207170_Z_17_Z)

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HDAC1 and 2 (HDAC1/2) form the catalytic core of four co-repressor complexes (Sin3A, NuRD, CoREST and MiDAC) which have essential roles in DNA replication, cell cycle progression and gene expression. The goal of the Cowley lab is to define a network of accessory proteins, target genes and substrates (including non-histones), for each of the four canonical HDAC1/2 complexes. In this application we intend to contribute to this process by identifying novel associated proteins and substrates of the prototypical HDAC1/2 complex, Sin3A, using a new technique termed, Biotin-ID (Bio-ID). Bio-ID uses a promiscuous version of the biotin ligase, BirA (BirA*), fused to the protein of interest, thus allowing ‘proximate’ proteins (within approx. 10nm) to be labelled with Biotin, which can then be identified using a streptavidin pull-down and mass-spectrometry. The advantage of this approach is that it allows the identification of both binding partners and transiently associated proteins such as substrates. A parallel study with the LSD1/CoREST complex in the Cowley lab has identified known binding partners (CoREST1-3, HDAC1/2, and BHC80), substrates (p53), as well as 23 other novel chromatin associated proteins. We would therefore like to extend this approach to the study of the Sin3A/HDAC1 complex.

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Grant Details

Amount Awarded 0
Applicant Surname Bailey
Approval Committee Internal Decision Panel
Award Date 2017-04-27T00:00:00+00:00
Financial Year 2016/17
Grant Programme: Title Vacation Scholarships
Internal ID 207170/Z/17/Z
Lead Applicant Mr Luke Bailey
Partnership Value 0
Planned Dates: End Date 2017-08-04T00:00:00+00:00
Planned Dates: Start Date 2017-06-05T00:00:00+00:00
Recipient Org: Country United Kingdom
Region East Midlands