Human PGBD5 DNA transposase promotes site-specific oncogenic mutations in rhabdoid tumors

Henssen, Anton; Koche, Richard P.; Zhuang, Jiali; Jiang, Eileen; Reed, Casie; Eisenberg, Amy; Still, Eric; MacArthur, Ian; Rodríguez-Fos, Elias; González, Santiago; Puiggròs, Montserrat; Blackford, Andrew N.; Mason, Christopher E.; Stanchina, Elisa de; Gönen, Mithat; Emde, Anne-Katrin; Shah, Minita; Arora, Kanika; Reeves, Catherine; Socci, Nicholas D.; Perlman, Elizabeth J.; Antonescu, Cristina R.; Roberts, Charles W.M.; Steen, Hanno; Mullen, Elizabeth; Jackson, Stephen; Torrents, David; Weng, Zhiping; Armstrong, Scott A.; Kentsis, Alex

doi:10.1101/111138

Cited by 3 publications

(20 citation statements)

References 45 publications

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“…3, RKRKKRK motif typed in green and underlined). In agreement with the literature [14] we observed that the ectopic expression of murine PGBD5 isoform of 523 amino acids (Mm523) reduced the apparent efficiency of obtaining NeoR clones (Fig. 5a) that is close to that of PB.NLS-1-558 in HeLa cells (Fig.…”

Section: Pgbd5 a Natural Domesticated Crd-deficient Pble Transposasesupporting

confidence: 92%

“…3c and 6d), but it displayed a marked difference in that there were fourfold and twofold less proper insertion events by transposition than in those obtained with PB.NLS-1-558 and Mm523, respectively. Unexpectedly, this suggests that PGBD5 is more prone to trigger improper integration in rhabdoid tumor G401 cells, consistent with the proposal that PGBD5 exhibits aberrant activities in human rhabdoid tumors [14]. Since we observed that PB.NLS-1-558, Mm523 and Hs524 display similar junction profile, we wondered if their insertion site preferences might be similar.…”

Section: Pgbd5 a Natural Domesticated Crd-deficient Pble Transposasesupporting

confidence: 82%

“…5b) was similar to that obtained with the GFP control. In order to verify whether this was due to PGBD5 cytotoxicity, we used a second cellular system developed in human rhabdoid tumor G401 cells and in which the endogenous expression of PGBD5 (Hs524) was found to have little impact on cell viability [14]. Under these experimental conditions, we found that the rate of NeoR clones was sevenfold higher than that of the GFP control (Fig.…”

Section: Pgbd5 a Natural Domesticated Crd-deficient Pble Transposasementioning

confidence: 94%

“…Overall, this means that PB.NLS-1-558 is roughly 8 times less efficient than PB for obtaining NeoR clones in an integration assay done in HeLa cells. Because cytotoxicity hampered our ability to directly evaluate integration rates and was likely dose-dependent as observed with PGBD5 [14], we focused our investigation on the ability of PB.NLS-1-558 to trigger Ifp2 transposition by characterizing integration events into chromosomes by NeoR clones.…”

Section: Pb Variants Display Cytotoxicitymentioning

confidence: 99%

“…4) revealed that both CRDdeficient proteins displayed an acidic N-terminal domain and a second domain with a basic pI (~9.2) containing an apparent catalytic triad composed of 3 acidic amino acid residues that were essential for transposition activity [20]. Another shared feature was their ability to trigger Ifp2 transposition [14,21]. This transposition ability was rather unexpected for PGBD5 compared to PB.NLS-1-558 because the PGDB5 catalytic triad was not located at the same positions as in pble transposases (Fig.…”

Section: Pgbd5 a Natural Domesticated Crd-deficient Pble Transposasementioning

confidence: 99%

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The C-terminal Domain of piggyBac Transposase Is Not Required for DNA Transposition

Helou

Dardente

Arensburger

et al. 2021

Journal of Molecular Biology

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Section: Pgbd5 a Natural Domesticated Crd-deficient Pble Transposasesupporting

confidence: 92%

Section: Pgbd5 a Natural Domesticated Crd-deficient Pble Transposasesupporting

confidence: 82%

Section: Pgbd5 a Natural Domesticated Crd-deficient Pble Transposasementioning

confidence: 94%

Section: Pb Variants Display Cytotoxicitymentioning

confidence: 99%

Section: Pgbd5 a Natural Domesticated Crd-deficient Pble Transposasementioning

confidence: 99%

See 3 more Smart Citations

The C-terminal Domain of piggyBac Transposase Is Not Required for DNA Transposition

Helou

Dardente

Arensburger

et al. 2021

Journal of Molecular Biology

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Analysis of DNA transposition by DNA transposases in human cells

Bigot

Yamada

Mueller

et al. 2023

Preprint

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This manuscript discusses the recent report "Cognate restriction of transposition by piggyBac-like proteins" in Nucleic Acids Research by Beckermann et al and related recent publications about the inability to detect DNA transposition activity of human domesticated DNA transposase PGBD5. Measuring DNA transposition activity of transposases in human cells, where these enzymes can act on endogenous genomic substrates and induce DNA damage, is complicated by these and other cellular responses. Possible reasons for the discordant findings of Beckermann et al with prior independent reports of PGBD5 DNA transposition by Helou et al and Henssen et al and specific details of experimental methods in human cells are presented. In particular, by using independent experiments that reproduce PGBD5-mediated genomic integration, we demonstrate how supraphysiologic and ectopic overexpression of PGBD5 can cause DNA damage and cell death, and artifactual loss of apparent activity in clonogenic transposition reporter assays. While PGBD5 can support apparent DNA transposition, its cellular activity predominantly involves double-strand DNA breaks, deletions and other DNA rearrangements. We discuss the implications of this phenomenon for the interpretation of experimental assays and activities of domesticated DNA transposases.

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The Pgbd5 DNA transposase is required for mouse cerebral cortex development through DNA double-strand breaks formation

Simi

Ansaloni

Damiani

et al. 2023

Preprint

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Transposons are conserved mobile genetic elements and one of the major sources of genetic variability during organisms evolution. PiggyBac Transposable Element Derived 5 (Pgbd5), an evolutionarily conserved vertebrate DNA transposase-coding gene, is highly expressed in the central nervous system during development and adult life, suggesting it may play a role in neuronal differentiation and maintenance. Here, we show that Pgbd5 controls the expression of pro-neuronal and proliferative genes in a cell autonomous manner, leading to a decreased generation of neurons with altered migration. This activity depends on the induction of endogenous DNA double-strand breaks (DSBs). These results position Pgbd5 as a crucial regulator of brain development through its endonuclease activity.

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Human PGBD5 DNA transposase promotes site-specific oncogenic mutations in rhabdoid tumors

Cited by 3 publications

References 45 publications

The C-terminal Domain of piggyBac Transposase Is Not Required for DNA Transposition

The C-terminal Domain of piggyBac Transposase Is Not Required for DNA Transposition

Analysis of DNA transposition by DNA transposases in human cells

The Pgbd5 DNA transposase is required for mouse cerebral cortex development through DNA double-strand breaks formation

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