PCNA has specific functions in regulation of metabolism in haematological cells

Røst, Lisa Marie; Olaisen, Camilla; Sharma, Animesh; Nedal, Aina; Petrović, Vesna; Kvitvang, Hans Fredrik Nyvold; Bruheim, Per; Otterlei, Marit

doi:10.1101/2020.04.29.067512

Cited by 4 publications

(5 citation statements)

References 55 publications

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“…Metabolite abbreviations and HMDB IDs are listed in Supplementary Table S2A. These results are previously published in BioRxiv [49]. cellular stress introduced by LPS, and not only by DNA damage, as shown previously [6], increased the affinity of APIM to PCNA.…”

Section: Discussionsupporting

confidence: 81%

PCNA regulates primary metabolism by scaffolding metabolic enzymes

et al. 2022

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The essential roles of proliferating cell nuclear antigen (PCNA) as a scaffold protein in DNA replication and repair are well established, while its cytosolic roles are less explored. Two metabolic enzymes, alpha-enolase (ENO1) and 6-phosphogluconate dehydrogenase (6PGD), both contain PCNA interacting motifs. Mutation of the PCNA interacting motif APIM in ENO1 (F423A) impaired its binding to PCNA and resulted in reduced cellular levels of ENO1 protein, reduced growth rate, reduced glucose consumption, and reduced activation of AKT. Metabolome and signalome analysis reveal large consequences of impairing the direct interaction between PCNA and ENO1. Metabolites above ENO1 in glycolysis accumulated while lower glycolytic and TCA cycle metabolite pools decreased in the APIM-mutated cells; however, their overall energetic status were similar to parental cells. Treating haematological cancer cells or activated primary monocytes with a PCNA targeting peptide drug containing APIM (ATX-101) also lead to a metabolic shift characterized by reduced glycolytic rate. In addition, we show that ATX-101 treatments reduced the ENO1 - PCNA interaction, the ENO1, GAPDH and 6PGD protein levels, as well as the 6PGD activity. Here we report for the first time that PCNA acts as a scaffold for metabolic enzymes, and thereby act as a direct regulator of primary metabolism.

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Section: Discussionsupporting

confidence: 81%

PCNA regulates primary metabolism by scaffolding metabolic enzymes

et al. 2022

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“…Especially noteworthy is the slight increase in pyrimidine nucleoside pools in stavudine single treated cells, as opposed to down-regulated pools in both in the APIM-peptide-and the combination-treated group. Down-regulation of nucleoside pools after treatment with APIM-peptide has also been also detected in mammalian cancer cells [52]. However, the increased mutation frequency observed by stavudine as a single agent cannot be explained by cellular stress induced by the small increase in nucleotide pools detected.…”

Section: Stavudine Treatment Induces Only Minor Changes In Central Ca...mentioning

confidence: 94%

“…peptide has also been also detected in mammalian cancer cells [52]. However, the increased mutation frequency observed by stavudine as a single agent cannot be explained by cellular stress induced by the small increase in nucleotide pools detected.…”

Section: Stavudine Treatment Induces Only Minor Changes In Central Ca...mentioning

confidence: 95%

Nucleoside Analogues Are Potent Inducers of Pol V-mediated Mutagenesis

et al. 2021

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Drugs targeting DNA and RNA in mammalian cells or viruses can also affect bacteria present in the host and thereby induce the bacterial SOS system. This has the potential to increase mutagenesis and the development of antimicrobial resistance (AMR). Here, we have examined nucleoside analogues (NAs) commonly used in anti-viral and anti-cancer therapies for potential effects on mutagenesis in Escherichia coli, using the rifampicin mutagenicity assay. To further explore the mode of action of the NAs, we applied E. coli deletion mutants, a peptide inhibiting Pol V (APIM-peptide) and metabolome and proteome analyses. Five out of the thirteen NAs examined, including three nucleoside reverse transcriptase inhibitors (NRTIs) and two anti-cancer drugs, increased the mutation frequency in E. coli by more than 25-fold at doses that were within reported plasma concentration range (Pl.CR), but that did not affect bacterial growth. We show that the SOS response is induced and that the increase in mutation frequency is mediated by the TLS polymerase Pol V. Quantitative mass spectrometry-based metabolite profiling did not reveal large changes in nucleoside phosphate or other central carbon metabolite pools, which suggests that the SOS induction is an effect of increased replicative stress. Our results suggest that NAs/NRTIs can contribute to the development of AMR and that drugs inhibiting Pol V can reverse this mutagenesis.

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“…These are important observations from the following perspectives too. Cytoplasmic PCNA has recently been shown to control glucose metabolism in hematological cells (31). Survivin has been shown to increase the stability of oxidative phosphorylation Complex II, which enhances mitochondrial respiration, and thereby cancer metabolism (32).…”

Section: Discussionmentioning

confidence: 99%

Genetic Manipulation of Sirtuin 3 Causes Alterations of Key Metabolic Regulators in Melanoma

et al. 2021

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The mitochondrial sirtuin SIRT3 plays key roles in cellular metabolism and energy production, which makes it an obvious target for the management of cancer, including melanoma. Previously, we have demonstrated that SIRT3 was constitutively upregulated in human melanoma and its inhibition resulted in anti-proliferative effects in vitro in human melanoma cells and in vivo in human melanoma xenografts. In this study, we expanded our data employing knockdown and overexpression strategies in cell culture and mouse xenografts to further validate and establish the pro-proliferative function of SIRT3 in melanocytic cells, and its associated potential mechanisms, especially focusing on the metabolic regulation. We found that short-hairpin RNA (shRNA) mediated SIRT3 knockdown in G361 melanoma cells showed diminished tumorigenesis in immunodeficient Nu/Nu mice. Conversely, SIRT3 overexpressing Hs294T melanoma cells showed increased tumor growth. These effects were consistent with changes in markers of proliferation (PCNA), survival (Survivin) and angiogenesis (VEGF) in xenografted tissues. Further, in in vitro culture system, we determined the effect of SIRT3 knockdown on glucose metabolism in SK-MEL-2 cells, using a PCR array. SIRT3 knockdown caused alterations in a total of 37 genes involved in the regulation and enzymatic pathways of glucose (32 genes) and glycogen (5 genes) metabolism. Functions annotation of these identified genes, using the ingenuity pathway analysis (IPA), predicted cumulative actions of decreased cell viability/proliferation, tumor growth and reactive oxygen species (ROS), and increased apoptosis in response to SIRT3 knockdown. Further, IPA gene network analysis of SIRT3 modulated genes revealed the interactions among these genes in addition to several melanoma-associated genes. Sirtuin pathway was identified as one of the top canonical pathways showing the interaction of SIRT3 with metabolic regulatory genes along with other sirtuins. IPA analysis also predicted the inhibition of HIF1α, PKM, KDM8, PPARGC1A, mTOR, and activation of P53 and CLPP; the genes involved in major cancer/melanoma-associated signaling events. Collectively, these results suggest that SIRT3 inhibition affects cellular metabolism, to impart an anti-proliferative response against melanoma.

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PCNA has specific functions in regulation of metabolism in haematological cells

Cited by 4 publications

References 55 publications

PCNA regulates primary metabolism by scaffolding metabolic enzymes

PCNA regulates primary metabolism by scaffolding metabolic enzymes

Nucleoside Analogues Are Potent Inducers of Pol V-mediated Mutagenesis

Genetic Manipulation of Sirtuin 3 Causes Alterations of Key Metabolic Regulators in Melanoma

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