Mechanism, regulation, and functional properties of Dictyostelium myosin-1B.
Myosin-1B is one of three long tailed class-1 myosins containing an ATP-insensitive actin-binding site in the tail region that are produced in Dictyostelium discoideum. Myosin-1B localizes to actin-rich structures at the leading edge of migrating cells where it has been implicated in the formation and retraction of membrane projections, the recycling of plasma membrane components, and intracellular particle transport. Here, we have used a combination of molecular engineering approaches to describe the kinetic and motile properties of the myosin-1B motor and its regulation by TEDS site phosphorylation. Our results show that myosin-1B is a low duty ratio motor and displays the fastest nucleotide binding kinetics of any of the Dictyostelium class-1 myosins studied so far. Different from Dictyostelium myosin-1D and myosin-1E, dephosphorylated myosin-1B is not inactivated but moves actin filaments efficiently, albeit at an up to 8-fold slower velocity in the in vitro motility assay. A further difference is that myosin-1B lacks the ability to switch between rapid movement and bearing tension upon physiological changes of free Mg2+ ions. In this respect, its motor properties appear to be more closely related to Dictyostelium myosin-2 and rabbit skeletal muscle myosin.
Tsiavaliaris
Georgios
G
Institute for Biophysical Chemistry, OE 4350, Hannover Medical School, Carl-Neuberg-Strasse 1, D-30623 Hannover, Germany. gtsiaval@bpc.mh-hannover.de
Fujita-Becker
Setsuko
S
Dürrwang
Ulrike
U
Diensthuber
Ralph P
RP
Geeves
Michael A
MA
Manstein
Dietmar J
DJ
D008841:Actin Cytoskeleton / Q000235:genetics / Q000378:metabolism*; D000199:Actins / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D001692:Biological Transport / Q000502:physiology; D022081:Cell Surface Extensions / Q000235:genetics / Q000378:metabolism*; D004023:Dictyostelium / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D008274:Magnesium / Q000378:metabolism; D009218:Myosins / Q000235:genetics / Q000378:metabolism*; D009711:Nucleotides / Q000378:metabolism; D011485:Protein Binding / Q000502:physiology; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D011817:Rabbits; D013045:Species Specificity
D000199:Actins; D009711:Nucleotides; D015800:Protozoan Proteins; D009218:Myosins; D008274:Magnesium