Retromer protein VPS29 plays a crucial and positive role in the sumoylation system mediated by E3 SUMO ligase SIZ1

Vacuolar protein sorting 29 (VPS29) functions in retrograde protein transport as a component of the retromer complex. However, the role of VPS29 in the regulation of post-translational modifications, such as sumoylation and ubiquitination, has not been elucidated. In this study, we demonstrate that VPS29 positively regulates SIZ/PIAS-type E3 SUMO (Small ubiquitin-related modifier) ligase-mediated sumoylation systems. In Arabidopsis, vps29-3 mutants display upregulated salicylic acid (SA) signaling pathways and reactive oxygen species accumulation, similar to those observed in siz1 mutants. Arabidopsis VPS29 (AtVPS29) directly interacts with the Arabidopsis E3 SUMO ligase SIZ1 (AtSIZ1) and localizes not only to the cytoplasm but also to the nucleus. The loss of AtVPS29 leads to a depletion of AtSIZ1, whereas the E3 ubiquitin ligase constitutive photomorphogenic 1 (COP1), an upstream regulator of AtSIZ1, accumulates in vps29-3 mutants. Conversely, overexpression of AtVPS29 results in the accumulation of AtSIZ1 and the depletion of COP1 in transgenic Arabidopsis. Similarly, in human cells, silencing of hVPS29 leads to the depletion of the E3 SUMO ligase, PIAS1, and the accumulation of huCOP1. Under heat stress conditions, the levels of SUMO-conjugates are significantly lower in Arabidopsis vps29-3 mutants, indicating a regulatory role of AtVPS29 on AtSIZ1 activity. Moreover, AtVPS29 inhibits ubiquitination pathway-dependent degradation of AtSIZ1. Notably, AtSIZ1 forms a complex with AtVPS29 and trimeric retromer proteins. Taken together, our results indicate that VPS29 plays an essential role in signal transduction by regulating SIZ/PIAS-type E3 ligase-dependent sumoylation in both plants and animals.