TY - JOUR
T1 - RNA ligase-like domain in activating signal cointegrator 1 complex subunit 1 (ASCC1) regulates ASCC complex function during alkylation damage
AU - Soll, Jennifer M.
AU - Brickner, Joshua R.
AU - Mudge, Miranda C.
AU - Mosammaparast, Nima
N1 - Funding Information:
This work was supported in part by National Institutes of Health Grant R01 CA193318, the Alvin Siteman Cancer Research Fund, the Siteman Investment Program, and the Children’s Discovery Institute of St. Louis Children’s Hospital Grant MC-II-2015-453 (to N. M.). The authors declare that they have no conflicts of interest with the contents of this article. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Funding Information:
Acknowledgments—We thank Hani Zaher and members of the Mosammaparast lab for their advice on this manuscript. We acknowledge the Alvin J. Siteman Cancer Center at Washington University and Barnes-Jewish Hospital for the use of the GEiC Core. The Siteman Cancer Center is supported by NCI Cancer Center Support Grant P30 CA091842 from the National Institutes of Health.
Funding Information:
This work was supported in part by National Institutes of Health Grant R01 CA193318, the Alvin Siteman Cancer Research Fund, the Siteman Invest-ment Program, and the Children’s Discovery Institute of St. Louis Children’s Hospital Grant MC-II-2015-453 (to N. M.). The authors declare that they have no conflicts of interest with the contents of this article. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Funding Information:
1 Supported by a graduate student fellowship from the Monsanto Company.
Publisher Copyright:
© 2018 by The American Society for Biochemistry and Molecular Biology, Inc.
PY - 2018/8/31
Y1 - 2018/8/31
N2 - Multiple DNA damage response (DDR) pathways have evolved to sense the presence of damage and recruit the proper repair factors. We recently reported a signaling pathway induced upon alkylation damage to recruit the AlkB homolog 3, -ketoglutarate– dependent dioxygenase (ALKBH3)–activating signal cointegrator 1 complex subunit 3 (ASCC3) dealkylase–helicase repair complex. As in other DDR pathways, the recruitment of these repair factors is mediated through a ubiquitin-de-pendent mechanism. However, the machinery that coordinates the proper assembly of this repair complex and controls its recruitment is still poorly defined. Here, we demonstrate that the ASCC1 accessory subunit is important for the regulation of ASCC complex function. ASCC1 interacts with the ASCC complex through the ASCC3 helicase subunit. We find that ASCC1 is present at nuclear speckle foci prior to damage, but leaves the foci in response to alkylation. Strikingly, ASCC1 loss significantly increases ASCC3 foci formation during alkylation damage, yet most of these foci lack ASCC2. These results suggest that ASCC1 coordinates the proper recruitment of the ASCC complex during alkylation, a function that appears to depend on a putative RNA-binding motif near the ASCC1 C terminus. Consistent with its role in alkylation damage signaling and repair, ASCC1 knockout through a CRISPR/Cas9 approach results in alkylation damage sensitivity in a manner epistatic with ASCC3. Together, our results identify a critical regulator of the ALKBH3–ASCC alkylation damage signaling pathway and suggest a potential role for RNA-interacting domains in the alkylation damage response.
AB - Multiple DNA damage response (DDR) pathways have evolved to sense the presence of damage and recruit the proper repair factors. We recently reported a signaling pathway induced upon alkylation damage to recruit the AlkB homolog 3, -ketoglutarate– dependent dioxygenase (ALKBH3)–activating signal cointegrator 1 complex subunit 3 (ASCC3) dealkylase–helicase repair complex. As in other DDR pathways, the recruitment of these repair factors is mediated through a ubiquitin-de-pendent mechanism. However, the machinery that coordinates the proper assembly of this repair complex and controls its recruitment is still poorly defined. Here, we demonstrate that the ASCC1 accessory subunit is important for the regulation of ASCC complex function. ASCC1 interacts with the ASCC complex through the ASCC3 helicase subunit. We find that ASCC1 is present at nuclear speckle foci prior to damage, but leaves the foci in response to alkylation. Strikingly, ASCC1 loss significantly increases ASCC3 foci formation during alkylation damage, yet most of these foci lack ASCC2. These results suggest that ASCC1 coordinates the proper recruitment of the ASCC complex during alkylation, a function that appears to depend on a putative RNA-binding motif near the ASCC1 C terminus. Consistent with its role in alkylation damage signaling and repair, ASCC1 knockout through a CRISPR/Cas9 approach results in alkylation damage sensitivity in a manner epistatic with ASCC3. Together, our results identify a critical regulator of the ALKBH3–ASCC alkylation damage signaling pathway and suggest a potential role for RNA-interacting domains in the alkylation damage response.
UR - http://www.scopus.com/inward/record.url?scp=85052550877&partnerID=8YFLogxK
U2 - 10.1074/jbc.RA117.000114
DO - 10.1074/jbc.RA117.000114
M3 - Article
C2 - 29997253
AN - SCOPUS:85052550877
VL - 293
SP - 13524
EP - 13533
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 35
ER -