Redox-dependent condensation and cytoplasmic granulation by human ssDNA binding protein 1 delineate roles in oxidative stress response

Harami, G. M.; Palinkas, J.; Kovacs, Z. J.; Tarnok, K.; Harami-Papp, H.; Hegedus, J.; Mahmudova, L.; Kucsma, N.; Toth, S.; Szakacs, G.; Kovacs, M.

doi:10.1101/2023.07.25.550517

bioRxiv · 10.1101/2023.07.25.550517

Redox-dependent condensation and cytoplasmic granulation by human ssDNA binding protein 1 delineate roles in oxidative stress response

Harami, G. M.·Palinkas, J.·Kovacs, Z. J.·Tarnok, K.·Harami-Papp, H.·Hegedus, J.·Mahmudova, L.·Kucsma, N.·Toth, S.·Szakacs, G.·Kovacs, M.

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Abstract

Human single-stranded DNA binding protein 1 (hSSB1/NABP2/OBFC2B) plays central roles in the repair of DNA breaks and oxidized DNA lesions. Here we show that hSSB1 undergoes liquid-liquid phase separation (LLPS) that is redox-dependent and requires the presence of single-stranded DNA or RNA, features that are distinct from those of LLPS by bacterial SSB. hSSB1 nucleoprotein droplets form under physiological ionic conditions, in response to treatment resulting in cellular oxidative stress. hSSB1s intrinsically disordered region (IDR) is indispensable for LLPS, whereas all three cysteine residues of the oligonucleotide/oligosaccharide-binding (OB) fold are necessary to maintain redox-sensitive droplet formation. Proteins interacting with hSSB1 show selective enrichment inside hSSB1 droplets, suggesting tight content control and recruitment functions for the condensates. While these features appear instrumental for genome repair, we also detected hSSB1 condensates in the cytoplasm in response to oxidative stress in various cell lines. hSSB1 condensates colocalize with stress granules, implying unexplored extranuclear roles in cellular stress response. Our results suggest novel, condensation-linked roles for hSSB1, linking genome repair and cytoplasmic defense.

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Harami, G. M., Palinkas, J., Kovacs, Z. J., Tarnok, K., Harami-Papp, H., Hegedus, J., Mahmudova, L., Kucsma, N., Toth, S., Szakacs, G., Kovacs, M.. 2023-07-25. Redox-dependent condensation and cytoplasmic granulation by human ssDNA binding protein 1 delineate roles in oxidative stress response. https://doi.org/10.1101/2023.07.25.550517

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