posted on 2021-12-16, 22:04authored byHugo Avila, Anh Truong, David Tyrpak, Shin-Jae Park, Siqi Lei, Yaocun Li, Curtis Okamoto, Sarah Hamm-Alvarez, J. Andrew MacKay
Dynamin (DNM) is a family of large
GTPases possessing a unique
mechanical ability to “pinch” off vesicles entering
cells. DNM2 is the most ubiquitously expressed member of the DNM family.
We developed a novel tool based on elastin-like polypeptide (ELP)
technology to quickly, precisely, and reversibly modulate the structure
of DNM2. ELPs are temperature-sensitive biopolymers that self-assemble
into microdomains above sharp transition temperatures. When linked
together, DNM2 and a temperature-sensitive ELP fusion organize into
a range of distinct temperature-dependent structures above a sharp
transition temperature, which were not observed with wild-type DNM2
or a temperature-insensitive ELP fusion control. The structures comprised
three different morphologies, which were prevalent at different temperature
ranges. The size of these structures was influenced by an inhibitor
of the DNM2 GTPase activity, dynasore; furthermore, they appear to
entrap co-expressed cytosolic ELPs. Having demonstrated an unexpected
diversity of morphologically distinct structures, DNM2–ELP
fusions may have applications in the exploration of dynamin-dependent
biology.