THE HERPESVIRUS NUCLEAR EGRESS COMPLEX COMPONENT, UL31, IS RECRUITED TO SITES OF DNA DAMAGE THROUGH POLY(ADP-RIBOSE) BINDING

The herpes simplex virus (HSV) UL31 gene encodes a conserved member of the herpesvirus nuclear egress complex that not only functions in the egress of DNA-containing capsids from the nucleus, but is also required for optimal viral genome expression, replication and packaging into capsids. Here, we report that the UL31 protein from HSV-2 and the orthologous protein, ORF69, from Kaposi's sarcoma-associated herpesvirus (KSHV) are recruited to sites of DNA damage. Recruitment of UL31 to sites of DNA damage occurred in HSV-2 infected cells, but did not require other viral proteins. The N-terminus of UL31 contains sequences resembling a poly(ADP-ribose) (PAR) binding motif. As protein poly-ADP ribosylation (PARylation) is a hallmark of the DNA damage response we examined the relationship between PARylation and UL31 recruitment to DNA damage. While the PAR polymerase (PARP)1/2 inhibitor, olaparib, prevented UL31 recruitment to damaged DNA, KU55933 inhibition of signaling through the ataxia telangiectasia mutated (ATM) DNA damage response pathway had no effect. These findings were further supported by experiments demonstrating direct and specific interaction between HSV-2 UL31 and PAR using purified components. Co-transfection with the viral kinase Us3, known to phosphorylate UL31, inhibited UL31 recruitment to DNA damage but also prevented the recruitment of other proteins recruited to DNA damage sites. The viral E3 ubiquitin ligase ICP0 was observed to co-localize with UL31 in transfected cells in a manner that is independent of the PAR-binding ability of UL31. However, inhibition of PARP1/2/3 did not reduce the ability of HSV-2 to replicate and we observed reduced PAR levels in the nuclei of infected cells. This study reveals a previously unrecognized function for UL31 orthologs and may suggest that the recognition of PAR by UL31 is coupled to the nuclear egress of herpesvirus capsids, influences viral DNA replication and packaging, or possibly modulates the DNA damage response mounted by virally infected cells.

RE-MEANING THE SACRED: COLONIAL DAMAGE AND INDIGENOUS COSMOLOGIES

Indigenous ways of knowing are dependent on an inheriting process both amongst humans and between human and non-human being. These multi-relationships cross material and immaterial borders as sites of knowledge production. This manuscript will interrogate how three particular Indigenous cosmological relationships have been purposefully re-meaninged by colonial institutions: 1) How Anishinaabe and Haudenosaunee origin stories have been abstracted into a distinctive epistemological versus ontological site; 2) How Anishnaabe spirit worlds are impacted by colonial relations, and how state institutions benefit from the re-meaning of these worlds; and 3) How Indigenous sovereignty in Canada is imagined from a statist perspective, and how these polices have re-meaninged the sacred relationships within a cosmological understanding of Haudenosaunee governance. The re-meaning of sacredly-held Indigenous relationships is both accelerated by, and contributes to, a practice of reducing upon Indigenous and non-human societies. Throughout expressions of colonialism on Indigenous territories (the academy, the state, Indian policy), Indigenous knowledge is consistently either dismissed or appropriated. This reduction of Indigenous knowledge continues to bolster functions of the state as related to the elimination of the “Indian Problem” via reducing the “Indian” to an adaptive subject.