title: Molecular basis of dynamic relocalization of Dictyostelium myosin IB.
abstract: Class I myosins have a single heavy chain comprising an N-terminal motor domain with actin-activated ATPase activity and a C-terminal globular tail with a basic region that binds to acidic phospholipids. These myosins contribute to the formation of actin-rich protrusions such as pseudopodia, but regulation of the dynamic localization to these structures is not understood. Previously, we found that Acanthamoeba myosin IC binds to acidic phospholipids in vitro through a short sequence of basic and hydrophobic amino acids, BH site, based on the charge density of the phospholipids. The tail of Dictyostelium myosin IB (DMIB) also contains a BH site. We now report that the BH site is essential for DMIB binding to the plasma membrane and describe the molecular basis of the dynamic relocalization of DMIB in live cells. Endogenous DMIB is localized uniformly on the plasma membrane of resting cells, at active protrusions and cell-cell contacts of randomly moving cells, and at the front of motile polarized cells. The BH site is required for association of DMIB with the plasma membrane at all stages where it colocalizes with phosphoinositide bisphosphate/phosphoinositide trisphosphate (PIP(2)/PIP(3)). The charge-based specificity of the BH site allows for in vivo specificity of DMIB for PIP(2)/PIP(3) similar to the PH domain-based specificity of other class I myosins. However, DMIB-head is required for relocalization of DMIB to the front of migrating cells. Motor activity is not essential, but the actin binding site in the head is important. Thus, dynamic relocalization of DMIB is determined principally by the local PIP(2)/PIP(3) concentration in the plasma membrane and cytoplasmic F-actin.
authors.affiliation: Laboratory of Cell Biology, NHLBI, National Institutes of Health, Bethesda, Maryland 20892, USA. brzeskah@mail.nih.gov
authors.forename: Hanna
authors.initials: H
authors.lastname: Brzeska
authors.forename: Jake
authors.initials: J
authors.lastname: Guag
authors.forename: G Michael
authors.initials: GM
authors.lastname: Preston
authors.forename: Margaret A
authors.initials: MA
authors.lastname: Titus
authors.forename: Edward D
authors.initials: ED
authors.lastname: Korn
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D002462:Cell Membrane / Q000235:genetics / Q000378:metabolism*; D004023:Dictyostelium / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D024461:Myosin Type I / Q000235:genetics / Q000378:metabolism*; D018129:Phosphatidylinositol Phosphates / Q000235:genetics / Q000378:metabolism; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D011554:Pseudopodia / Q000235:genetics / Q000378:metabolism*
chemical_list: D000199:Actins; D018129:Phosphatidylinositol Phosphates; D015800:Protozoan Proteins; C060974:phosphatidylinositol 3,4,5-triphosphate; C060980:phosphatidylinositol 3,4-diphosphate; D024461:Myosin Type I