Analysis of the role of nucleotides in axonemal dynein function

Axonemal dynein in flagella and cilia is a motor molecule that produces microtubule sliding, powered by the energy of ATP hydrolysis. Our goal is to understand how dynein motile activity is controlled to produce the characteristic oscillatory movement of flagella. ATP, the energy source for dynein, is also important as a regulator of dynein activity. Among the four nucleotide-binding sites of a dynein heavy chain, one is the primary ATP hydrolyzing site while the others are noncatalytic sites and thought to perform regulatory functions. Stable binding of both ATP and ADP to these regulatory sites is probably essential for the chemomechanical energy transduction in dynein. Although the ATP concentration in beating flagella is physiologically high and constant, at any moment in the oscillatory cycle some dynein molecules are active while others are not, and the motile activity of dynein oscillates temporally and spatially in the axoneme. It is likely that the basic mechanism underlying the highly dynamic control of dynein activity involves the ATP-dependent inhibition and ADP-dependent activation (or release of inhibition) of dynein. How the inhibition and activation can be induced in beating flagella is still unknown. It seems, however, that the mechanical force of bending is involved in the activation of dynein, probably through the control of noncatalytic nucleotide binding to dynein. This chapter provides an overview of several approaches, using sea urchin sperm flagella, to studying the roles of ATP and ADP in the regulation of dynein activity with or without the mechanical signal of bending.