Histone H1.2 Dependent Translocation of Poly (ADP-ribose) Initiates Parthanatos

Abstract

Toxic cellular insults activate the nuclear protein poly (ADP-ribose) (PAR) polymerase-1 (PARP-1) to initiate parthanatos, a regulated cell death program. PAR acts as a death signal by translocating from the nucleus to the cytosol, where it activates the next steps in the parthanatic cell death cascade. How PAR translocates from the nucleus to the cytosol is not known. Here we show that PARylation and PAR binding to histone H1.2 enables it to act as a carrier, transporting PAR out of the nucleus to the cytosol. Knocking down the expression of histone H1.2 via CRISPR/Cas9 and knockout of histone H1.2 reduces the translocation of PAR to the cytosol after treatment of human cortical neurons with N-methyl-D-aspartate (NMDA) or following oxygen-glucose deprivation (OGD). The PAR-dependent E3 ubiquitin ligase, Iduna (RNF146) ubiquitinates PARylated H1.2. Overexpression of Iduna reduces the expression levels of cytosolic histone H1.2, preventing the translocation of PAR following NMDA or OGD exposure, similar to inhibition of PAR formation by the PARP inhibitor, DPQ. Whereas, the catalytically null variant Iduna C60A, or the PAR binding mutant Iduna Y156A and R157A (YRAA) was ineffective in ubiquitinating histone H1.2 and preventing the reduction in cytosolic histone H1.2 levels and PAR translocation from the nucleus to the cytosol. Histone H1.2 heterozygote and homozygote knockout mice exhibited reduced infarct volume 24 hrs post middle cerebral artery occlusion (MCAO) and showed better recovery in motor deficits than wildtype littermates at day 3 and/or day 7 post MCAO. Collectively, these findings reveal histone H1.2 as the key carrier of PAR out of the nucleus to the cytosol where it participates in the next step of the parthanatic cell death cascade.