Family affairs: ties and bonds among protein phosphatases with Kelch-like domains in plants

GUSTAVO A. MASELLI; SANTIAGO MORA-GARCIA

Turku

Congreso; Europhosphatase 2015: Phosphorylation switches and cellular homeostasis; 2015

EMBO

Protein phosphatases with Kelch-like domains (PPKL) are members of the PPP family present in green algae, plants and alveolates. PPKL are bimodular proteins with a β-propeller domain at the N-terminus and a catalytic domain akin to PP1 at the C-terminus; both domains are tethered by an intermediate sequence of unknown affiliations. In plants, PPKLs have been described as effectors of brassinosteroid signaling and cell proliferation. We reappraised the roles of the four members of the family present in Arabidopsis thaliana -BSL1, BSL2, BSL3 and BSU1- through phylogenetic, functional and genetic analyses. BSL1 and BSL2/BSL3 belong to two ancient evolutionary clades that have been highly conserved in land plants; in contrast, BSU1-type genes are exclusively found in the Brassicaceae and display a remarkable sequence divergence, even among closely related species. Simultaneous loss of function of the closely related paralogs BSL2 and BSL3 causes a peculiar array of phenotypic alterations; loss of function of BSL1 is, in turn, phenotypically silent. However, the three major paralogs play together an essential role in early stages of development, that BSU1 is unable to supplement. These evolutionary patterns mirror an unusual behaviour in PPP phosphatases: the ability of PPKL to form homotypic interactions. BSL1, BSL2 and BSL3 behave in vivo as homodimers, whereas BSU1 is only able of transient interactions. The interaction determinants are, however, different among members of the main clades. In BSL1, the catalytic domain is the main driver of interaction; in contrast, the catalytic domain of BSL2 is necessary but not sufficient, and dimerization requires two conserved sequence stretches in the linker domain. The linker domain of BSL2 behaves in solution as an intrinsically disordered protein prone to form secondary structure elements upon mild perturbations. Several functional and regulatory layers thus appear to be at work in this small and essential gene family.