コンテンツへスキップ
Merck
  • The human Shu complex functions with PDS5B and SPIDR to promote homologous recombination.

The human Shu complex functions with PDS5B and SPIDR to promote homologous recombination.

Nucleic acids research (2019-10-31)
Julieta Martino, Gregory J Brunette, Jonathan Barroso-González, Tatiana N Moiseeva, Chelsea M Smith, Christopher J Bakkenist, Roderick J O'Sullivan, Kara A Bernstein
要旨

RAD51 plays a central role in homologous recombination during double-strand break repair and in replication fork dynamics. Misregulation of RAD51 is associated with genetic instability and cancer. RAD51 is regulated by many accessory proteins including the highly conserved Shu complex. Here, we report the function of the human Shu complex during replication to regulate RAD51 recruitment to DNA repair foci and, secondly, during replication fork restart following replication fork stalling. Deletion of the Shu complex members, SWS1 and SWSAP1, using CRISPR/Cas9, renders cells specifically sensitive to the replication fork stalling and collapse caused by methyl methanesulfonate and mitomycin C exposure, a delayed and reduced RAD51 response, and fewer sister chromatid exchanges. Our additional analysis identified SPIDR and PDS5B as novel Shu complex interacting partners and genetically function in the same pathway upon DNA damage. Collectively, our study uncovers a protein complex, which consists of SWS1, SWSAP1, SPIDR and PDS5B, involved in DNA repair and provides insight into Shu complex function and composition.

材料
製品番号
ブランド
製品内容

Millipore
抗FLAG® M2抗体 アフィニティーゲル, purified immunoglobulin, buffered aqueous glycerol solution
Sigma-Aldrich
モノクローナル抗FLAG® M2抗体 マウス宿主抗体, clone M2, purified immunoglobulin (Purified IgG1 subclass), buffered aqueous solution (10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide)
Millipore
エンドヌクレアーゼ Serratia marcescens(セラチア菌)由来
Sigma-Aldrich
マイトマイシンC Streptomyces caespitosus由来, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
メタンスルホン酸メチル, 99%
Sigma-Aldrich
ヒドロキシ尿素, 98%, powder
Sigma-Aldrich
cis-ジアンミン白金(II) ジクロリド, crystalline
Sigma-Aldrich
フェニルメチルスルホニルフルオリド, ≥99.0% (T)
Sigma-Aldrich
5-ブロモ-2′-デオキシウリジン, BioUltra, ≥99%
Sigma-Aldrich
デメコルシン 溶液, 10 μg/mL in HBSS, ACF Qualified, BioXtra
Sigma-Aldrich
ビオチン, ≥99% (HPLC), lyophilized powder
Sigma-Aldrich
ビスベンズイミドH 33258, powder, BioReagent, suitable for cell culture, ≥98% (HPLC and TLC)
Sigma-Aldrich
マグネシウムメチルカルボナート 溶液, 2.0 M in DMF
Supelco
メタンスルホン酸メチル, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Sigma-Aldrich
Anti-SPIDR antibody produced in rabbit, affinity isolated antibody