Abstract
FANCA is a key player in the canonical Fanconi anemia (FA) repair pathway. We have recently shown that FANCA also plays an important role in the single-strand annealing sub-pathway (SSA) of DNA double-strand break (DSB) repair by biochemically catalyzing single-strand annealing. Here, we report that a steroidal lactone withaferin A (WA) specifically impedes SSA repair by promoting FANCA downregulation at a sub-micromolar concentration range. We find that WA causes FANCA downregulation post-translationally in a proteasome-dependent manner. This WA-mediated downregulation is achieved through HSP90 inhibition and disruption of the FANCA-HSP90 interaction. WA-mediated FANCA degradation significantly reduces cellular SSA repair, abolishes FANCD2 monoubiquitination, elevates sensitivity to mitomycin C, and results in accumulation of DSBs. Importantly, the WA-induced defect in SSA repair is highly dependent on the absence of FANCA protein and overexpression of exogenous WT-FANCA protein in WA-treated cells significantly complements the repair defect.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 10-17 |
| Number of pages | 8 |
| Journal | DNA Repair |
| Volume | 77 |
| DOIs | |
| State | Published - May 1 2019 |
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Keywords
- Double strand break repair
- FANCA
- Fanconi anemia
- Single strand annealing
ASJC Scopus subject areas
- Biochemistry
- Molecular Biology
- Cell Biology
Cite this
Impeding the single-strand annealing pathway of DNA double-strand break repair by withaferin A-mediated FANCA degradation. / Liu, Wenjun; Wang, Guanying; Palovcak, Anna; Li, Yan; Hao, Sophie; Liu, Zhao Jun; Landgraf, Ralf; Yuan, Fenghua; Zhang, Yanbin.
In: DNA Repair, Vol. 77, 01.05.2019, p. 10-17.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Impeding the single-strand annealing pathway of DNA double-strand break repair by withaferin A-mediated FANCA degradation
AU - Liu, Wenjun
AU - Wang, Guanying
AU - Palovcak, Anna
AU - Li, Yan
AU - Hao, Sophie
AU - Liu, Zhao Jun
AU - Landgraf, Ralf
AU - Yuan, Fenghua
AU - Zhang, Yanbin
PY - 2019/5/1
Y1 - 2019/5/1
N2 - FANCA is a key player in the canonical Fanconi anemia (FA) repair pathway. We have recently shown that FANCA also plays an important role in the single-strand annealing sub-pathway (SSA) of DNA double-strand break (DSB) repair by biochemically catalyzing single-strand annealing. Here, we report that a steroidal lactone withaferin A (WA) specifically impedes SSA repair by promoting FANCA downregulation at a sub-micromolar concentration range. We find that WA causes FANCA downregulation post-translationally in a proteasome-dependent manner. This WA-mediated downregulation is achieved through HSP90 inhibition and disruption of the FANCA-HSP90 interaction. WA-mediated FANCA degradation significantly reduces cellular SSA repair, abolishes FANCD2 monoubiquitination, elevates sensitivity to mitomycin C, and results in accumulation of DSBs. Importantly, the WA-induced defect in SSA repair is highly dependent on the absence of FANCA protein and overexpression of exogenous WT-FANCA protein in WA-treated cells significantly complements the repair defect.
AB - FANCA is a key player in the canonical Fanconi anemia (FA) repair pathway. We have recently shown that FANCA also plays an important role in the single-strand annealing sub-pathway (SSA) of DNA double-strand break (DSB) repair by biochemically catalyzing single-strand annealing. Here, we report that a steroidal lactone withaferin A (WA) specifically impedes SSA repair by promoting FANCA downregulation at a sub-micromolar concentration range. We find that WA causes FANCA downregulation post-translationally in a proteasome-dependent manner. This WA-mediated downregulation is achieved through HSP90 inhibition and disruption of the FANCA-HSP90 interaction. WA-mediated FANCA degradation significantly reduces cellular SSA repair, abolishes FANCD2 monoubiquitination, elevates sensitivity to mitomycin C, and results in accumulation of DSBs. Importantly, the WA-induced defect in SSA repair is highly dependent on the absence of FANCA protein and overexpression of exogenous WT-FANCA protein in WA-treated cells significantly complements the repair defect.
KW - Double strand break repair
KW - FANCA
KW - Fanconi anemia
KW - Single strand annealing
UR - http://www.scopus.com/inward/record.url?scp=85062219564&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85062219564&partnerID=8YFLogxK
U2 - 10.1016/j.dnarep.2019.02.010
DO - 10.1016/j.dnarep.2019.02.010
M3 - Article
C2 - 30844655
AN - SCOPUS:85062219564
VL - 77
SP - 10
EP - 17
JO - DNA Repair
JF - DNA Repair
SN - 1568-7864
ER -