TY - JOUR
T1 - Extended DNA threading through a dual-engine motor module of the activating signal co-integrator 1 complex
AU - Jia, Junqiao
AU - Hilal, Tarek
AU - Bohnsack, Katherine E.
AU - Chernev, Aleksandar
AU - Tsao, Ning
AU - Bethmann, Juliane
AU - Arumugam, Aruna
AU - Parmely, Lane
AU - Holton, Nicole
AU - Loll, Bernhard
AU - Mosammaparast, Nima
AU - Bohnsack, Markus T.
AU - Urlaub, Henning
AU - Wahl, Markus C.
N1 - Funding Information:
We thank Agnieszka Pietrzyk-Brzezinska (Lodz University of Technology, Poland) for help in the cloning of the ALKBH3 expression construct, Philipp Hackert (University Medical Center Göttingen, Germany) for technical assistance, Ralf Pflanz and Monika Raabe (Max-Planck-Institut für Multidisziplinäre Naturwissenschaften, Germany) for help in mass spectrometric analysis and Daniel Lauster (Freie Universität Berlin, Germany) for advice in MST analyses. We acknowledge the assistance of the core facility BioSupraMol supported by the Deutsche Forschungsgemeinschaft in electron microscopic analyses. This work was supported by grants from the Deutsche Forschungsgemeinschaft (INST 130/1064−1 FUGG to Freie Universität Berlin; SFB1565 [project number 469281184], to K.E.B., M.T.B., H.U., and M.C.W.; BO 3442/1-2 [project number 192916677] to M.T.B.; SFB860 [project number 105286809] to K.E.B. and H.U.), the American Cancer Society (RSG−18−156-01-DMC to N.M.), the National Institutes of Health of the U.S. (R01 CA193318 and P01 CA092584 to N.M.) and the Berlin University Alliance (501_BIS-CryoFac to M.C.W.).
Funding Information:
We thank Agnieszka Pietrzyk-Brzezinska (Lodz University of Technology, Poland) for help in the cloning of the ALKBH3 expression construct, Philipp Hackert (University Medical Center Göttingen, Germany) for technical assistance, Ralf Pflanz and Monika Raabe (Max-Planck-Institut für Multidisziplinäre Naturwissenschaften, Germany) for help in mass spectrometric analysis and Daniel Lauster (Freie Universität Berlin, Germany) for advice in MST analyses. We acknowledge the assistance of the core facility BioSupraMol supported by the Deutsche Forschungsgemeinschaft in electron microscopic analyses. This work was supported by grants from the Deutsche Forschungsgemeinschaft (INST 130/1064−1 FUGG to Freie Universität Berlin; SFB1565 [project number 469281184], to K.E.B., M.T.B., H.U., and M.C.W.; BO 3442/1-2 [project number 192916677] to M.T.B.; SFB860 [project number 105286809] to K.E.B. and H.U.), the American Cancer Society (RSG−18−156-01-DMC to N.M.), the National Institutes of Health of the U.S. (R01 CA193318 and P01 CA092584 to N.M.) and the Berlin University Alliance (501_BIS-CryoFac to M.C.W.).
Publisher Copyright:
© 2023, The Author(s).
PY - 2023/12
Y1 - 2023/12
N2 - Activating signal co-integrator 1 complex (ASCC) subunit 3 (ASCC3) supports diverse genome maintenance and gene expression processes, and contains tandem Ski2-like NTPase/helicase cassettes crucial for these functions. Presently, the molecular mechanisms underlying ASCC3 helicase activity and regulation remain unresolved. We present cryogenic electron microscopy, DNA-protein cross-linking/mass spectrometry as well as in vitro and cellular functional analyses of the ASCC3-TRIP4 sub-module of ASCC. Unlike the related spliceosomal SNRNP200 RNA helicase, ASCC3 can thread substrates through both helicase cassettes. TRIP4 docks on ASCC3 via a zinc finger domain and stimulates the helicase by positioning an ASC-1 homology domain next to the C-terminal helicase cassette of ASCC3, likely supporting substrate engagement and assisting the DNA exit. TRIP4 binds ASCC3 mutually exclusively with the DNA/RNA dealkylase, ALKBH3, directing ASCC3 for specific processes. Our findings define ASCC3-TRIP4 as a tunable motor module of ASCC that encompasses two cooperating NTPase/helicase units functionally expanded by TRIP4.
AB - Activating signal co-integrator 1 complex (ASCC) subunit 3 (ASCC3) supports diverse genome maintenance and gene expression processes, and contains tandem Ski2-like NTPase/helicase cassettes crucial for these functions. Presently, the molecular mechanisms underlying ASCC3 helicase activity and regulation remain unresolved. We present cryogenic electron microscopy, DNA-protein cross-linking/mass spectrometry as well as in vitro and cellular functional analyses of the ASCC3-TRIP4 sub-module of ASCC. Unlike the related spliceosomal SNRNP200 RNA helicase, ASCC3 can thread substrates through both helicase cassettes. TRIP4 docks on ASCC3 via a zinc finger domain and stimulates the helicase by positioning an ASC-1 homology domain next to the C-terminal helicase cassette of ASCC3, likely supporting substrate engagement and assisting the DNA exit. TRIP4 binds ASCC3 mutually exclusively with the DNA/RNA dealkylase, ALKBH3, directing ASCC3 for specific processes. Our findings define ASCC3-TRIP4 as a tunable motor module of ASCC that encompasses two cooperating NTPase/helicase units functionally expanded by TRIP4.
UR - http://www.scopus.com/inward/record.url?scp=85151783735&partnerID=8YFLogxK
U2 - 10.1038/s41467-023-37528-3
DO - 10.1038/s41467-023-37528-3
M3 - Article
C2 - 37019967
AN - SCOPUS:85151783735
SN - 2041-1723
VL - 14
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 1886
ER -