Conformational alteration of DOCK5•ELMO1 signalosome on lipid membrane

TL;DR

This study reveals how the DOCK5•ELMO1 complex changes shape on cell membranes to regulate signaling, using cryo-EM and biochemical experiments.

Contribution

A new conformation of the DOCK5•ELMO1 complex on lipid membranes is identified, showing how it regulates GEF activity.

Findings

DOCK5•ELMO1 adopts a flattened, planar structure on lipid membranes via hinge rotations.

Acidic lipids drive conformational changes that regulate DOCK5 GEF activity and downstream signaling.

The study provides a method to analyze large lipid-associated complexes like signalosomes.

Abstract

The DOCK protein family activates Rho small GTPases through guanine nucleotide exchange factor (GEF) activity. DOCK is thought to exert its GEF activity at the plasma membrane. However, the mechanism by which DOCK activity on the plasma membrane is regulated remains unclear. Herein, we present a new conformation in which DOCK5, ELMO1, RhoG, and Rac1 are aligned on a plane and symmetrically flattened, as revealed by cryo-EM using a lipid membrane-coated grid. The major conformational change leading to this structure results from rotation of each DOCK5•ELMO1 hinge site through interactions with the membrane. Biochemical and cellular experiments indicate that conformational changes driven by acidic lipids are important for regulating the GEF activity of the DOCK5•ELMO1 complex on the plasma membrane and are essential for its downstream signalling. This approach also enables the analysis of large lipid-associated complexes, such as signalosomes, and will aid studies of membrane-dependent signalling assemblies.

Membrane-flattened DOCK5–ELMO1–RhoG–Rac1 complexes reveal that conformational changes from hinge rotations and ELMO1 PH domain–lipid interactions cooperatively regulate DOCK5 GEF activity and downstream signalling.

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