コンテンツへスキップ
Merck

ATRX proximal protein associations boast roles beyond histone deposition.

PLoS genetics (2021-11-16)
William A Scott, Erum Z Dhanji, Boris J A Dyakov, Ema S Dreseris, Jonathon S Asa, Laura J Grange, Mila Mirceta, Christopher E Pearson, Grant S Stewart, Anne-Claude Gingras, Eric I Campos
要旨

The ATRX ATP-dependent chromatin remodelling/helicase protein associates with the DAXX histone chaperone to deposit histone H3.3 over repetitive DNA regions. Because ATRX-protein interactions impart functions, such as histone deposition, we used proximity-dependent biotinylation (BioID) to identify proximal associations for ATRX. The proteomic screen captured known interactors, such as DAXX, NBS1, and PML, but also identified a range of new associating proteins. To gauge the scope of their roles, we examined three novel ATRX-associating proteins that likely differed in function, and for which little data were available. We found CCDC71 to associate with ATRX, but also HP1 and NAP1, suggesting a role in chromatin maintenance. Contrastingly, FAM207A associated with proteins involved in ribosome biosynthesis and localized to the nucleolus. ATRX proximal associations with the SLF2 DNA damage response factor help inhibit telomere exchanges. We further screened for the proteomic changes at telomeres when ATRX, SLF2, or both proteins were deleted. The loss caused important changes in the abundance of chromatin remodelling, DNA replication, and DNA repair factors at telomeres. Interestingly, several of these have previously been implicated in alternative lengthening of telomeres. Altogether, this study expands the repertoire of ATRX-associating proteins and functions.

材料
製品番号
ブランド
製品内容

Sigma-Aldrich
プロテアーゼインヒビターカクテル, for use with mammalian cell and tissue extracts, DMSO solution
Sigma-Aldrich
モノクローナル抗FLAG® M2抗体 マウス宿主抗体, 1 mg/mL, clone M2, affinity isolated antibody, buffered aqueous solution (50% glycerol, 10 mM sodium phosphate, and 150 mM NaCl, pH 7.4)
Sigma-Aldrich
ポリ-L-リシン 溶液, 0.1 % (w/v) in H2O
Sigma-Aldrich
DAPI, for nucleic acid staining
Millipore
エンドヌクレアーゼ Serratia marcescens(セラチア菌)由来
Sigma-Aldrich
Duolink® In Situ検出試薬FarRed
Roche
ブロッキング試薬, For nucleic acid hybridization and detection
Sigma-Aldrich
タージトール 溶液, Type NP-40, 70% in H2O
Sigma-Aldrich
ビスベンズイミドH 33258, for fluorescence, ≥98.0% (HPLC)
Sigma-Aldrich
抗ZFHX4抗体 ウサギ宿主抗体, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution
Sigma-Aldrich
Anti-EZH2 Antibody, clone AE25, clone AE25, from mouse