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bioRxiv · 10.1101/2025.08.07.669170

Reverse transcriptase inhibitors induce autophagy in a LINE-1 ORF1p-dependent manner

Abstract

Human Long Interspersed Nuclear Element-1 (LINE-1) retrotransposons propagate throughout the genome via reverse-transcribed RNA intermediates. LINE-1 expression is pervasive in cancer. Functional LINE-1s encode two proteins: ORF1p, an RNA-binding protein, and ORF2p, harboring reverse transcriptase and endonuclease activities. Reverse transcriptase inhibitors, including non-nucleoside (NNRTI) and nucleoside (NRTI) inhibitors, inhibit cancer cell proliferation and antagonize cancer progression. We previously found that two NNRTIs induced DNA damage, nuclear lamin rupture, micronuclei formation, and autophagy in prostate cancer cells. We now find that two different RTIs up-regulate LINE-1 mRNA expression and ORF1p abundance in nuclei, triggering ORF1p interactions with lamin B1 and with DNA damage factors. ORF1p accumulates within micronuclei with damaged DNA and with the autophagy receptor p62. We further demonstrate that inhibiting autophagy, or decreasing ORF1p levels, prevent DNA damage and preserve lamin B1 integrity, uncoverig a role of LINE-1-ORF1p in the autophagy response of cancer cells, independent on retrotranscription events.

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BibTeXRIS

Damizia, M., Baranzini, M., Sciamanna, I., Altieri, L., Rovella, P., Bringa, O. G. R., Hozeifi, S., Saba, L. J., Mourtzinos, A., OShea, A., Ansari, M. S. Z., Andreoli, F., Cirilli, R., Sbardella, G., Serafino, A., Schumann, G. G., Trisciuoglio, D., LaCava, J., Lavia, P., Spadafora, C.. 2025-08-11. Reverse transcriptase inhibitors induce autophagy in a LINE-1 ORF1p-dependent manner. https://doi.org/10.1101/2025.08.07.669170

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