TY - JOUR
T1 - Structural basis of DNA synthesis opposite 8-oxoguanine by human PrimPol primase-polymerase
AU - Rechkoblit, Olga
AU - Johnson, Robert E.
AU - Gupta, Yogesh K.
AU - Prakash, Louise
AU - Prakash, Satya
AU - Aggarwal, Aneel K.
N1 - Funding Information:
We thank the staff at Northeastern Collaborative Access Team (NECAT) beam line at the Argonne National Laboratory for facilitating X-ray data collection. This work was partially supported by NIH grants R35-GM131780 (A. K. A.) and R01 GM126087 (L. P.). The NECAT beamline was funded by the National Institute of General Medical Sciences (NIGMS) from the NIH (P41 GM103403). The Pilatus 6 M detector on 24-ID-C beamline was funded by an NIH–Office of Research Infrastructure Programs High-End Instrumentation grant (S10 RR029205). This research used resources of the Advanced Photon Source, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under contract no. DE-AC02-06CH11357. Y. K. G is supported by the Max and Minnie Tomerlin Voelcker Foundation and the Cancer Prevention Research Institute of Texas (RP190534).
Publisher Copyright:
© 2021, The Author(s).
PY - 2021/12/1
Y1 - 2021/12/1
N2 - PrimPol is a human DNA polymerase-primase that localizes to mitochondria and nucleus and bypasses the major oxidative lesion 7,8-dihydro-8-oxoguanine (oxoG) via translesion synthesis, in mostly error-free manner. We present structures of PrimPol insertion complexes with a DNA template-primer and correct dCTP or erroneous dATP opposite the lesion, as well as extension complexes with C or A as a 3′−terminal primer base. We show that during the insertion of C and extension from it, the active site is unperturbed, reflecting the readiness of PrimPol to accommodate oxoG(anti). The misinsertion of A opposite oxoG(syn) also does not alter the active site, and is likely less favorable due to lower thermodynamic stability of the oxoG(syn)•A base-pair. During the extension step, oxoG(syn) induces an opening of its base-pair with A or misalignment of the 3′-A primer terminus. Together, the structures show how PrimPol accurately synthesizes DNA opposite oxidatively damaged DNA in human cells.
AB - PrimPol is a human DNA polymerase-primase that localizes to mitochondria and nucleus and bypasses the major oxidative lesion 7,8-dihydro-8-oxoguanine (oxoG) via translesion synthesis, in mostly error-free manner. We present structures of PrimPol insertion complexes with a DNA template-primer and correct dCTP or erroneous dATP opposite the lesion, as well as extension complexes with C or A as a 3′−terminal primer base. We show that during the insertion of C and extension from it, the active site is unperturbed, reflecting the readiness of PrimPol to accommodate oxoG(anti). The misinsertion of A opposite oxoG(syn) also does not alter the active site, and is likely less favorable due to lower thermodynamic stability of the oxoG(syn)•A base-pair. During the extension step, oxoG(syn) induces an opening of its base-pair with A or misalignment of the 3′-A primer terminus. Together, the structures show how PrimPol accurately synthesizes DNA opposite oxidatively damaged DNA in human cells.
UR - http://www.scopus.com/inward/record.url?scp=85109048809&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85109048809&partnerID=8YFLogxK
U2 - 10.1038/s41467-021-24317-z
DO - 10.1038/s41467-021-24317-z
M3 - Article
C2 - 34188055
AN - SCOPUS:85109048809
SN - 2041-1723
VL - 12
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 4020
ER -