Biomedical subjects

Shigehiko Yumura

Publications and source records attributed to Shigehiko Yumura.

7 recordsLinked to original sources

PubMed2005

Multiple myosin II heavy chain kinases: roles in filament assembly control and proper cytokinesis in Dictyostelium.

Myosin II filament assembly in Dictyostelium discoideum is regulated via phosphorylation of residues located in the carboxyl-terminal portion of the myosin II heavy chain (MHC) tail. A series of novel protein kinases in this system are capable of phosphorylating these residues in vitro, driving filament disassembly. Previous studies have demonstrated that at least three of these kinases (MHCK A, MHCK B, and MHCK C) display differential localization patterns in living cells. We have created a collection of single, double, and triple gene knockout cell lines for this family of kinases. Analysis of these lines reveals that three MHC kinases appear to represent the majority of cellular activity capable of driving myosin II filament disassembly, and reveals that cytokinesis defects increase with the number of kinases disrupted. Using biochemical fractionation of cytoskeletons and in vivo measurements via fluorescence recovery after photobleaching (FRAP), we find that myosin II overassembly increases incrementally in the mutants, with the MHCK A(-)/B(-)/C(-) triple mutant showing severe myosin II overassembly. These studies suggest that the full complement of MHC kinases that significantly contribute to growth phase and cytokinesis myosin II disassembly in this organism has now been identified.

Animals↗

PubMed2005

De novo formation of basal bodies in Naegleria gruberi: regulation by phosphorylation.

The de novo formation of basal bodies in Naegleria gruberi was preceded by the transient formation of a microtubule (MT)-nucleating complex containing gamma-tubulin, pericentrin, and myosin II complex (GPM complex). The MT-nucleating activity of GPM complexes was maximal just before the formation of visible basal bodies and then rapidly decreased. The regulation of MT-nucleating activity of GPM complexes was accomplished by a transient phosphorylation of the complex. Inhibition of dephosphorylation after the formation of basal bodies resulted in the formation of multiple flagella. 2D-gel electrophoresis and Western blotting showed a parallel relationship between the MT-nucleating activity of GPM complexes and the presence of hyperphosphorylated gamma-tubulin in the complexes. These data suggest that the nucleation of MTs by GPM complexes precedes the de novo formation of basal bodies and that the regulation of MT-nucleating activity of GPM complexes is essential to the regulation of basal body number.

Animals↗

PubMed2004

Dynamics of novel feet of Dictyostelium cells during migration.

We observed the dynamics of actin foci in live Dictyostelium cells expressing GFP-actin. Actin foci were dynamic structures, but they were fixed on the substratum during cell migration. Interference reflection microscopy revealed that the ventral cell membrane was closer to the substratum at sites of actin foci. Furthermore, some actin foci were incorporated into the retraction fibers, ripped off from the cells and eventually shed on the substratum after the cells moved away. The velocity of the cells was inversely proportional to the number of actin foci. Measurement of traction force using a silicone substratum demonstrated that the traction force was transmitted to the substratum through actin foci. Taken together, several lines of evidence strongly suggest that actin foci function as the active 'feet' of Dictyostelium cells. We also found evidence suggesting that changing step is regulated in a coordinated manner during cell migration. Possible mechanisms by which these cells migrate across substrata are discussed in this context.

Actins↗

PubMed2004

Multiple parallelisms in animal cytokinesis.

The process of cytokinesis in animal cells is usually presented as a relatively simple picture: A cleavage plane is first positioned in the equatorial region by the astral microtubules of the anaphase mitotic apparatus, and a contractile ring made up of parallel filaments of actin and myosin II is formed and encircles the cortex at the division site. Active sliding between the two filament systems constricts the perimeter of the cortex, leading to separation of two daughter cells. However, recent studies in both animal cells and lower eukaryotic model organisms have demonstrated that cytokinesis is actually far more complex. It is now obvious that the three key processes of cytokinesis, cleavage plane determination, equatorial furrowing, and scission, are driven by different mechanisms in different types of cells. In some cases, moreover, multiple pathways appear to have redundant functions in a single cell type. In this review, we present a novel hypothesis that incorporates recent observations on the activities of mitotic microtubules and the biochemistry of Rho-type GTPase proteins and postulates that two different sets of microtubules are responsible for the two known mechanisms of cleavage plane determination and also for two distinct mechanisms of equatorial furrowing.

Actin Cytoskeleton↗

PubMed2003

Myosin II contributes to the posterior contraction and the anterior extension during the retraction phase in migrating Dictyostelium cells.

Cells must exert force against the substrate to migrate. We examined the vectors (both the direction and the magnitude) of the traction force generated by Dictyostelium cells using an improved non-wrinkling silicone substrate. During migration, the cells showed two 'alternate' phases of locomotory behavior, an extension phase and a retraction phase. In accordance with these phases, two alternate patterns were identified in the traction force. During the extension phase, the cell exerted a 'pulling force' toward the cell body in the anterior and the posterior regions and a 'pushing force' in the side of the cell (pattern 1). During the retraction phase, the cell exerted a 'pushing force' in the anterior region, although the force disappeared in the side and the posterior regions of the cell (pattern 2). Myosin II heavy chain null cells showed a single pattern in their traction force comparable to 'pattern 1', although they still had the alternate biphasic locomotory behavior similar to the wild-type cells. Therefore, the generation of 'pushing force' in the anterior and the cancellation of the traction force in the side and the posterior during the retraction phase were deficient in myosin knock-out mutant cells, suggesting that these activities depend on myosin II via the posterior contraction. Considering all these results, we hypothesized that there is a highly coordinated, biphasic mechanism of cell migration in Dictyostelium.

Animals↗

PubMed2003

Myosins and cell dynamics in cellular slime molds.

Myosin is a mechanochemical transducer and serves as a motor for various motile activities such as cell migration, cytokinesis, maintenance of cell shape, phagocytosis, and morphogenesis. Nonmuscle myosin in vivo does not either stay static at specific subcellular regions or construct highly organized structures, such as sarcomere in skeletal muscle cells. The cellular slime mold Dictyostelium discoideum is an ideal "model organism" for the investigation of cell movement and cytokinesis. The advantages of this organism prompted researchers to carry out pioneering cell biological, biochemical, and molecular genetic studies on myosin II, which resulted in elucidation of many fundamental features of function and regulation of this most abundant molecular motor. Furthermore, recent molecular biological research has revealed that many unconventional myosins play various functions in vivo. In this article, how myosins are organized and regulated in a dynamic manner in Dictyostelium cells is reviewed and discussed.

Animals↗

PubMed2002

Novel myosin heavy chain kinase involved in disassembly of myosin II filaments and efficient cleavage in mitotic dictyostelium cells.

We have cloned a full-length cDNA encoding a novel myosin II heavy chain kinase (mhckC) from Dictyostelium. Like other members of the myosin heavy chain kinase family, the mhckC gene product, MHCK C, has a kinase domain in its N-terminal half and six WD repeats in the C-terminal half. GFP-MHCK C fusion protein localized to the cortex of interphase cells, to the cleavage furrow of mitotic cells, and to the posterior of migrating cells. These distributions of GFP-MHCK C always corresponded with that of myosin II filaments and were not observed in myosin II-null cells, where GFP-MHCK C was diffusely distributed in the cytoplasm. Thus, localization of MHCK C seems to be myosin II-dependent. Cells lacking the mhckC gene exhibited excessive aggregation of myosin II filaments in the cleavage furrows and in the posteriors of the daughter cells once cleavage was complete. The cleavage process of these cells took longer than that of wild-type cells. Taken together, these findings suggest MHCK C drives the disassembly of myosin II filaments for efficient cytokinesis and recycling of myosin II that occurs during cytokinesis.

Amino Acid Motifs↗