TY - JOUR
T1 - A unique arginine cluster in PolDIP2 enhances nucleotide binding and DNA synthesis by PrimPol
AU - Kasho, Kazutoshi
AU - Stojkovič, Gorazd
AU - Velázquez-Ruiz, Cristina
AU - Martínez-Jiménez, Maria Isabel
AU - Doimo, Mara
AU - Laurent, Timothée
AU - Berner, Andreas
AU - Pérez-Rivera, Aldo E.
AU - Jenninger, Louise
AU - Blanco, Luis
AU - Wanrooij, Sjoerd
N1 - Funding Information:
Knut and Alice Wallenberg Foundation (to S.W.); Swedish Research Council (to S.W.); JSPS Overseas Research Fellowship [201860287 to K.K.]; Spanish Ministry of Economy and Competitiveness [BFU2015-65880-P and PGC2018-093576-B-C21 to L.B.]; C.V.R. was recipient of a FPU-predoctoral fellowship from Spanish Ministry of Economy and Competitiveness; G.S. was supported by the Kempe Foundation. Funding for open access charge: Knut och Alice Wallenbergs Stiftelse.
Publisher Copyright:
© 2021 The Author(s) 2021.
PY - 2021/2/26
Y1 - 2021/2/26
N2 - Replication forks often stall at damaged DNA. To overcome these obstructions and complete the DNA duplication in a timely fashion, replication can be restarted downstream of the DNA lesion. In mammalian cells, this repriming of replication can be achieved through the activities of primase and polymerase PrimPol. PrimPol is stimulated in DNA synthesis through interaction with PolDIP2, however the exact mechanism of this PolDIP2-dependent stimulation is still unclear. Here, we show that PrimPol uses a flexible loop to interact with the C-terminal ApaG-like domain of PolDIP2, and that this contact is essential for PrimPol's enhanced processivity. PolDIP2 increases primer-template and dNTP binding affinities of PrimPol, which concomitantly enhances its nucleotide incorporation efficiency. This stimulation is dependent on a unique arginine cluster in PolDIP2. Since the polymerase activity of PrimPol alone is very limited, this mechanism, where the affinity for dNTPs gets increased by PolDIP2 binding, might be critical for the in vivo function of PrimPol in tolerating DNA lesions at physiological nucleotide concentrations.
AB - Replication forks often stall at damaged DNA. To overcome these obstructions and complete the DNA duplication in a timely fashion, replication can be restarted downstream of the DNA lesion. In mammalian cells, this repriming of replication can be achieved through the activities of primase and polymerase PrimPol. PrimPol is stimulated in DNA synthesis through interaction with PolDIP2, however the exact mechanism of this PolDIP2-dependent stimulation is still unclear. Here, we show that PrimPol uses a flexible loop to interact with the C-terminal ApaG-like domain of PolDIP2, and that this contact is essential for PrimPol's enhanced processivity. PolDIP2 increases primer-template and dNTP binding affinities of PrimPol, which concomitantly enhances its nucleotide incorporation efficiency. This stimulation is dependent on a unique arginine cluster in PolDIP2. Since the polymerase activity of PrimPol alone is very limited, this mechanism, where the affinity for dNTPs gets increased by PolDIP2 binding, might be critical for the in vivo function of PrimPol in tolerating DNA lesions at physiological nucleotide concentrations.
UR - http://www.scopus.com/inward/record.url?scp=85102403658&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85102403658&partnerID=8YFLogxK
U2 - 10.1093/nar/gkab049
DO - 10.1093/nar/gkab049
M3 - Article
C2 - 33533925
AN - SCOPUS:85102403658
SN - 0305-1048
VL - 49
SP - 2179
EP - 2191
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 4
ER -