The plastoglobule-localized metallopeptidase PGM48 is a positive regulator of senescence in arabidopsis thaliana

Abstract

Plastoglobuli (PG) are thylakoid-associated monolayer lipid particles with a specific proteome of ~30 PG core proteins and isoprenoid and neutral lipids. During senescence, PGs increase in size, reflecting their role in dismantling thylakoid membranes. Here, we show that the only PG-localized peptidase PGM48 positively regulates leaf senescence. We discovered that PGM48 isa member of the M48 peptidase family with PGM48 homologs, forming a clade (M48D) only found in photosynthetic organisms.Unlike the M48A, B, and C clades, members of M48D have no transmembrane domains, consistent with their unique subcellular location in the PG. In vitro assays showed Zn-dependent proteolytic activity and substrate cleavage upstream of hydrophobic residues. Over expression of PGM48 accelerated natural leaf senescence, whereas suppression delayed senescence. Quantitative proteomics of PG from senescing rosettes of PGM48 over expression lines showed a dramatically reduced level of CAROTENOIDCLEAVAGE ENZYME4 (CCD4) and significantly increased levels of the senescence-induced ABC1 KINASE7 (ABC1K7) and PHYTYL ESTER SYNTHASE1 (PES1). Yeast two-hybrid experiments identified PG core proteins ABC1K3, PES1, and CCD4as PGM48 interactors, whereas several other PG-localized proteins and chlorophyll degradation enzymes did not interact. We discuss mechanisms through which PGM48 could possibly accelerate the senescence process.