Actin filament dynamics govern many key physiological processes from cell motility to tissue morphogenesis. A central feature of actin dynamics is the capacity of filaments to polymerize and depolymerize at their ends in response to cellular conditions. It is currently thought that filament kinetics can be described by a single rate constant for each end. In this study, using direct visualization of single actin filament elongation, we show that actin polymerization kinetics at both filament ends are strongly influenced by the binding of proteins to the lateral filament surface. We also show that the pointed-end has a non-elongating state that dominates the observed filament kinetic asymmetry. Estimates of flexibility as well as effects on fragmentation and growth suggest that the observed kinetic diversity arises from structural alteration. Tuning elongation kinetics by exploiting the malleability of the filament structure may be a ubiquitous mechanism to generate a rich variety of cellular actin dynamics.
Actin is one of the most abundant proteins in cells. It forms networks of filaments that provide structural support and generate the forces needed for cell movement, division, and many other processes in cells.
Filaments of actin continuously change in length as actin molecules are added or removed at the ends. One end of an actin filament—called the barbed-end—grows much faster than the other, known as the pointed-end. Many other proteins also help the actin filaments to form. Some of these proteins bind to the ends of the filaments, where they directly control the growth of the filaments. Other proteins bind along the length of the filaments, but how these ‘side-binding’ proteins influence the growth of filaments is not clear.
In this study, Crevenna et al. used a technique called ‘total internal reflection fluorescence (TIRF) microscopy’ to study how several side-binding proteins affect the growth of actin filaments in an artificial system. The growth of the barbed-ends was strongly influenced by which side-binding protein was interacting with the filament. For example, the barbed-end grew rapidly when a protein called VASP was present but grew more slowly in the presence of the protein α-actinin. Although the growth at the pointed-end was generally slow and sporadic, the side-binding proteins also had noticeable effects.
Crevenna et al. found that when the side-binding proteins were present at low levels, filament growth was similar for all proteins studied. It was only when the proteins were present at higher levels that the growth of the actin filaments was altered depending on the specific side-binding protein present. One side-binding protein called α-actinin also altered the shape of the actin filament so that when it was present at high levels, the filaments curved in a particular direction. Together, these results suggest that the growth, structure, and flexibility of actin filaments can be strongly influenced by the various proteins that bind along the length of the filaments.
The next challenges are to understand the precise details of how these side-binding proteins are able to alter the growth and shape of actin and investigate how they influence other processes that control the structure of actin networks in cells.