Small-molecule G-quadruplex stabilizers reveal a novel pathway of autophagy regulation in neurons

Moruno-Manchon, Jose F.; Lejault, Pauline; Wang, Yaoxuan; McCauley, Brenna S.; Honarpisheh, Pedram; Scheihing, Diego A Morales; Singh, Shivani; Dang, Weiwei; Kim, Nayun; Urayama, Akihiko; Zhu, Liang; Monchaud, David; McCullough, Louise D.; Tsvetkov, Andrey S.

doi:10.7554/elife.52283

Cited by 72 publications

(81 citation statements)

References 89 publications

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“…92 Pif1 has also been overexpressed in cells to reverse G4-mediated genetic instability in S. cerevisiae, 247 repair dysfunctions in cancer cells 248 and cognitive dysregulations in neurons. 249 In the latter two examples, Pif1 overexpression was found to remedy phenotypes of G4-mediated functional impairment induced by G4-stabilising PDS treatments. Similarly, BLM was overexpressed to reverse reactive oxygen species (ROS)-induced G4 (and R-loop) formation at transcriptionally active sites, thus restoring the repair process.…”

Section: The Dna Damage Response (Ddr)mentioning

confidence: 99%

DNA folds threaten genetic stability and can be leveraged for chemotherapy

et al. 2021

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Section: The Dna Damage Response (Ddr)mentioning

confidence: 99%

DNA folds threaten genetic stability and can be leveraged for chemotherapy

et al. 2021

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“…We recently investigated whether pharmacologically stabilizing G4-DNA with pyridostatin (PDS) [34], a selective G4-binding small molecule (referred to as a G4-ligand [35]), affects neuronal autophagy. Using circular dichroism spectroscopy (CD), thermal difference spectra (TDS) and nuclear magnetic resonance (NMR) analyses, we demonstrated that a putative G4-DNA forming sequence (PQFS) identified in the Atg7 gene folds into a stable G4 structure in vitro, which PDS strongly binds to [33]. We next showed that the G4-specific antibody (HF2) and the G4-binding protein SUB1/PC4 bind to the Atg7's PQFS ( Fig.…”

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confidence: 99%

“…The Atg7 gene (human, mouse, rat) contains numerous putative G4-forming motifs [33]. We recently investigated whether pharmacologically stabilizing G4-DNA with pyridostatin (PDS) [34], a selective G4-binding small molecule (referred to as a G4-ligand [35]), affects neuronal autophagy.…”

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Regulation of autophagy by DNA G-quadruplexes

Lejault¹,

Moruno-Manchon²,

Vemu³

et al. 2020

Autophagy

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Guanine-rich DNA strands can form secondary structures known as Gquadruplexes (G4-DNA). G4-DNA is important for the regulation of replication and transcription. We recently showed that the expression of Atg7, a gene that is critical for macroautophagy/autophagy, is controlled by G4-DNA in neurons. We demonstrated that the transcription factor SUB1/PC4 and the G4-DNA-specific antibody HF2 bind to a putative G4-DNA motif located in the Atg7 gene. Stabilizing G4-DNA with the G4-ligand pyridostatin (PDS) downregulates Atg7 expression in neurons. Here, we further investigated how G4-DNA in the Atg7 gene is stabilized by PDS. We show that PDS can form 1:1 and 2:1 complexes with the Atg7's G4. We also demonstrate that PDS downregulates the ATG7 protein and the expression of Atg7 in astrocytes as well as in neurons. Together with our previous findings, these data establish a novel G4-DNA-associated mechanism of autophagy regulation at a transcriptional level in neurons and astrocytes.

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“…The study of human PIF1 (hPIF1) has also demonstrated the deleterious consequences of Pif1 helicase misregulation. The transfection of cultured primary neurons with hPIF1 is toxic, increasing the risk of cell death (80). Further, human tumor cells rely on hPIF1 for protection from apoptosis (81), whereas hPIF1 depletion in normal cells increases replication fork arrest (82).…”

Section: Discussionmentioning

confidence: 99%

Lysine acetylation regulates the activity of nuclear Pif1

Ononye

Sausen

Balakrishnan

et al. 2020

Journal of Biological Chemistry

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In S. cerevisiae, the Pif1 helicase functions in both nuclear and mitochondrial DNA replication and repair processes, preferentially unwinding RNA-DNA hybrids and resolving G-quadruplex structures. We sought to determine how the various activities of Pif1 are regulated in vivo. Here, we report lysine acetylation of nuclear Pif1 and demonstrate that it influences both Pif1’s cellular roles and core biochemical activities. Using Pif1 overexpression toxicity assays, we determined that the acetyltransferase NuA4 and deacetylase Rpd3 are primarily responsible for the dynamic acetylation of nuclear Pif1. Mass spectrometry analysis revealed that Pif1 was modified in several domains throughout the protein’s sequence on the N-terminus (K118, K129), helicase domain (K525, K639, K725), and C-terminus (K800). Acetylation of Pif1 exacerbated its overexpression toxicity phenotype, which was alleviated upon deletion of its N-terminus. Biochemical assays demonstrated that acetylation of Pif1 stimulated its helicase, ATPase, and DNA binding activities while maintaining its substrate preferences. Limited proteolysis assays indicate that acetylation of Pif1 induces a conformational change that may account for its altered enzymatic properties. We propose that acetylation is involved in regulating of Pif1 activities, influencing a multitude of DNA transactions vital to the maintenance of genome integrity.

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Small-molecule G-quadruplex stabilizers reveal a novel pathway of autophagy regulation in neurons

Cited by 72 publications

References 89 publications

DNA folds threaten genetic stability and can be leveraged for chemotherapy

DNA folds threaten genetic stability and can be leveraged for chemotherapy

Regulation of autophagy by DNA G-quadruplexes

Lysine acetylation regulates the activity of nuclear Pif1

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