PolDIP2 interacts with human PrimPol and enhances its DNA polymerase activities

Guilliam, Thomas A.; Bailey, Laura J.; Brissett, Nigel C.; Doherty, Aidan J.

doi:10.1093/nar/gkw175

Cited by 60 publications

(102 citation statements)

References 32 publications

(109 reference statements)

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“…Poldip2 is fairly ubiquitously expressed, having been reported in aorta, 3, 4 lung, 3 kidney, 3, 5, 6 heart 7 and thyroid, 8 as well as many cell types including mouse aortic smooth muscle cells, 9 astrocytes, 10 vascular smooth muscle cells (VSMC), 3, 11 brain endothelial cells, 12 epithelial cells, 12 HeLa, 1, 12-15 fibroblasts, 2, 13, 16, 17 kidney fibroblasts, 18 myoblasts, 19 human umbilical vein endothelial cells, 20 human embryonic kidney cells 293 (HEK 293), 13 MRC5 cells, 21, 22 and cortical neurons. 23 There is also evidence of Poldip2 expression in human breast cancer, 24 human breast adenocarcinoma cells 13 and human lung adenocarcinoma cells.…”

Section: Tissue Expressionmentioning

confidence: 99%

“…In addition, Poldip2 stimulates the efficiency and error-free 8-oxo-7,8-dihydroguanine (8-oxo-G)DNA lesion bypass by PrimPol. 21 However, another study suggested that Poldip2 does not directly participate in the process of TLS, because it is not recruited to sites of DNA damage upon UV stimulation. Instead, Poldip2 translocates to spliceosomes and contributes to UV-induced MDM2 alternative splicing.…”

Section: Functions Of Poldip2mentioning

confidence: 99%

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Polymerase δ-interacting Protein 2: A Multifunctional Protein

Hernandes

Lassègue

Griendling

2017

Journal of Cardiovascular Pharmacology

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Polymerase δ-interacting protein 2 (Poldip2) is a multi-functional protein originally described as a binding partner of the p50 subunit of DNA polymerase δ and proliferating cell nuclear antigen. In addition to its role in DNA replication and damage repair, Poldip2 has been implicated in mitochondrial function, extracellular matrix regulation, cell cycle progression, focal adhesion turnover and cell migration. However, Poldip2 functions are incompletely understood. In this review, we discuss recent literature on Poldip2 tissue distribution, subcellular localization, and function. We also address the putative function of Poldip2 in cardiovascular disease, neurodegenerative conditions and in renal pathophysiology.

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Section: Tissue Expressionmentioning

confidence: 99%

Section: Functions Of Poldip2mentioning

confidence: 99%

Polymerase δ-interacting Protein 2: A Multifunctional Protein

Hernandes

Lassègue

Griendling

2017

Journal of Cardiovascular Pharmacology

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“…A number of studies have shown that PrimPol is important for the maintenance of replication after damage and loss of the protein causes UV-C sensitivity, slowing of replication and cell cycle arrest in avian DT40 cells (10,12,15,19). PrimPol has been shown to interact with a number of replicationassociated proteins, such as RPA and PolDIP2, which are likely to be important for its recruitment and function at sites of stalled replication (20)(21)(22).…”

Section: Introductionmentioning

confidence: 99%

PrimPol is required for the maintenance of efficient nuclear and mitochondrial DNA replication in human cells

Bailey

Bianchi

Doherty

2018

Preprint

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Eukaryotic Primase-Polymerase (PrimPol) is an enzyme that maintains efficient DNA duplication by repriming replication restart downstream of replicase stalling lesions and structures. To elucidate the cellular requirements for PrimPol in human cells, we generated PrimPol-deleted cell lines and show that it plays key roles in maintaining active replication in both the nucleus and mitochondrion, even in the absence of exogenous damage. Human cells lacking PrimPol exhibit delayed recovery after UV-C damage and increased mutation rates, micronuclei and sister chromatin exchanges but are not sensitive to genotoxins. PrimPol is also required during mitochondrial replication, with PrimPol-deficient cells having increased mtDNA copy number but displaying a significant decrease in replication rates. Deletion of PrimPol in XPV cells, lacking functional Pol Eta, causes an increase in DNA damage sensitivity and pronounced fork stalling after UV-C treatment. We show that, unlike canonical TLS polymerases, PrimPol is important for allowing active replication to proceed, even in the absence of exogenous damage, thus preventing the accumulation of excessive fork stalling and genetic mutations. Together, these findings highlight the importance of PrimPol for maintaining efficient DNA replication in unperturbed cells and its complementary roles, with Pol Eta, in damage tolerance in human cells.(200 words)

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“…An earlier study showed that human PolDIP2 increases the DNA polymerase activity of PrimPol (22). The crosslinking experiments in that study identified several regions as potential PrimPol-PolDIP2 interaction sites: however, the mechanism behind the PolDIP2-dependent stimulation of PrimPol DNA synthesis remained unclear.…”

Section: The C-terminal Region Of Poldip2 Increases the Processivity mentioning

confidence: 97%

A unique arginine cluster in PolDIP2 enhances nucleotide binding and DNA synthesis by PrimPol

Kasho

Stojkovič

Velázquez-Ruiz

et al. 2020

Preprint

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Replication forks often stall at damaged DNA. Resumption of DNA synthesis can occur by replacement of the replicative DNA polymerase with specialized, error-prone translesion DNA polymerases (TLS), that have higher tolerance for damaged substrates. Several of these polymerases (Polλ, Polη and PrimPol) are stimulated in DNA synthesis through interaction with PolDIP2, however the 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 PrimPol's primer-template and dNTP binding affinity, which concomitantly enhances PrimPol's nucleotide incorporation efficiency. This activity is dependent on a unique arginine cluster in PolDIP2 and could be essential for PrimPol to function in vivo, since the polymerase activity of PrimPol alone is very limited. This mechanism, where the affinity for dNTPs gets increased by PolDIP2 binding, could be common to all other PolDIP2interacting TLS polymerases, i.e. Polλ, Polη, Polζ and REV1, and might be critical for their in vivo function of tolerating DNA lesions at physiological nucleotide concentrations.replication fork progression (4-6). Among various TLS polymerases, the primase/polymerase PrimPol has been shown to help both in nuclear and mitochondrial DNA replication fork progression (7-8). Additionally, PrimPol can also re-prime downstream of blocking lesions, thus re-reinitiating DNA replication (1,(9)(10)(11)(12)(13)(14)(15).PrimPol is a monomeric enzyme belonging to the archaeal-eukaryotic primase (AEP) superfamily (16). Its catalytic core contains three highly conserved motifs (see Figure1A) which build the dNTP binding site used for elongation (17). To start primer synthesis, PrimPol requires a unique zinc finger (ZnF)-containing C-terminal domain, which facilitates binding of the first 5´-nucleotide (1, 18). Beside its function as a primase, PrimPol behaves in vitro as a polymerase with very low processivity (9, 19), suggesting the need for a cofactor to function optimally in the cell. In contrast to some other TLS polymerases (20), PrimPol's activity is not regulated by the sliding clamp Proliferating Cell Nuclear Antigen (PCNA) (21), suggesting that another factor might control its function. Indeed, polymerase d-interacting protein 2 (PolDIP2; also known as PDIP38) and single stranded DNA binding protein Replication Protein A (RPA) (22-24) are able to stimulate PrimPol-dependent DNA synthesis. Interestingly, PolDIP2was previously shown to physically interact with TLS polymerases Polz and REV1 (25), as well as to increase the processivity of Poll and Polh (26,27). Therefore, many TLS DNA polymerases could be regulated by a common, still to be elucidated, PolDIP2dependent mechanism.Here we show that PrimPol uses a flexible loop located at its AEP catalytic core to interact with the conserved ApaG-like C-terminal region of PolDIP2 (28), which is essential for the stimul...

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PolDIP2 interacts with human PrimPol and enhances its DNA polymerase activities

Cited by 60 publications

References 32 publications

Polymerase δ-interacting Protein 2: A Multifunctional Protein

Polymerase δ-interacting Protein 2: A Multifunctional Protein

PrimPol is required for the maintenance of efficient nuclear and mitochondrial DNA replication in human cells

A unique arginine cluster in PolDIP2 enhances nucleotide binding and DNA synthesis by PrimPol

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