Fluid-Phase Markers in the Basolateral Endocytic Pathway Accumulate in Response to the Actin Assembly-promoting Drug Jasplakinolide

Shurety, Wenda; Stewart, Nancy L.; Stow, Jennifer L.

doi:10.1091/mbc.9.4.957

Cited by 100 publications

(90 citation statements)

References 51 publications

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“…As we observed, high concentrations of jasplakinolide have been reported to increase the density of actin filaments adjacent the plasma membrane in both smooth muscle and Madin-Darby canine kidney cells (25,26), and this is accompanied by the loss of stress fibers in smooth muscle cells (26). Our description of changes in cellular architecture of fibroblasts after jasplakinolide treatment is consistent with those seen in epithelial cells (3).…”

Section: Discussionmentioning

confidence: 65%

Effects of Jasplakinolide on the Kinetics of Actin Polymerization

Bubb

Spector

Beyer

et al. 2000

Journal of Biological Chemistry

485

399

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Jasplakinolide paradoxically stabilizes actin filaments in vitro, but in vivo it can disrupt actin filaments and induce polymerization of monomeric actin into amorphous masses. A detailed analysis of the effects of jasplakinolide on the kinetics of actin polymerization suggests a resolution to this paradox. Jasplakinolide markedly enhances the rate of actin filament nucleation. This increase corresponds to a change in the size of actin oligomer capable of nucleating filament growth from four to approximately three subunits, which is mechanistically consistent with the localization of the jasplakinolide-binding site at an interface of three actin subunits. Because jasplakinolide both decreases the amount of sequestered actin (by lowering the critical concentration of actin) and augments nucleation, the enhancement of polymerization by jasplakinolide is amplified in the presence of actin-monomer sequestering proteins such as thymosin ␤ 4 . Overall, the kinetic parameters in vitro define the mechanism by which jasplakinolide induces polymerization of monomeric actin in vivo. Expected consequences of jasplakinolide function are consistent with the experimental observations and include de novo nucleation resulting in disordered polymeric actin and in insufficient monomeric actin to allow for remodeling of stress fibers.Jasplakinolide is a cyclic peptide isolated from the marine sponge, Jaspis johnstoni, that we have previously shown to bind to and stabilize filamentous actin in vitro (1). In vivo data suggests that jasplakinolide-treated prostate cancer cells have both decreased labeling of F-actin and decreased amounts of rhodamine-phalloidin bound to cell extracts (2), results that could be explained by the observation that jasplakinolide and phalloidin bind competitively to actin (1). In addition, however, in vivo data also convincingly show that jasplakinolide disrupts actin filaments with alterations in cellular architecture (2, 3), an effect that cannot be explained simply by competitive binding. We now present kinetic data characterizing the steady state and time-dependent in vitro interactions between jasplakinolide and actin that provide a plausible explanation for the effects of jasplakinolide on actin distribution in cultured cells. EXPERIMENTAL PROCEDURESMaterials-Rabbit skeletal muscle actin was prepared from frozen muscle (Pel-Freez, Rogers, AR) in buffer G (5.0 mM Tris, 0.2 mM ATP, 0.2 mM dithiothreitol, 0.1 mM CaCl 2 , and 0.01% sodium azide, pH 7.8) (4). Non-muscle actin from bovine brain was prepared by the method of Ruscha and Himes (5). Muscle and non-muscle pyrenyl-labeled actins 1 were prepared with 0.67-0.95 mol of label/mol of protein using the method of Kouyama and Mihashi (6). Labeled and unlabeled actins were further purified by gel filtration on Superose 12 (Amersham Pharmacia Biotech). Thymosin ␤ 4 cDNA was a gift from Dr. Vivian Nachmias and was inserted in a pET-11a vector, expressed in BL21(DE3) Escherichia coli, and purified as described previously (7). Jasplakinolide was a gift from ...

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Section: Discussionmentioning

confidence: 65%

Effects of Jasplakinolide on the Kinetics of Actin Polymerization

Bubb

Spector

Beyer

et al. 2000

Journal of Biological Chemistry

485

399

View full text Add to dashboard Cite

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“…n, number of PE analyzed. was described in other cell types, including in T. gondii (Bubb et al, 2000;Shurety et al, 1998;Wetzel et al, 2003). Recently, Smythe et al also observed by immunofluorescence microscopy that treatment of PE with 1 μM JAS for 5 hours caused a cortical redistribution of actin to the parasite periphery (Smythe et al, 2007).…”

Section: The Effects Of Jas and CD On Actin Distribution And Localizamentioning

confidence: 84%

A new model for hemoglobin ingestion and transport by the human malaria parasite Plasmodium falciparum

Lazarus

Schneider

Taraschi

2008

Journal of Cell Science

108

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The current model for hemoglobin ingestion and transport by intraerythrocytic Plasmodium falciparum malaria parasites shares similarities with endocytosis. However, the model is largely hypothetical, and the mechanisms responsible for the ingestion and transport of host cell hemoglobin to the lysosome-like food vacuole (FV) of the parasite are poorly understood. Because actin dynamics play key roles in vesicle formation and transport in endocytosis, we used the actin-perturbing agents jasplakinolide and cytochalasin D to investigate the role of parasite actin in hemoglobin ingestion and transport to the FV. In addition, we tested the current hemoglobin trafficking model through extensive analysis of serial thin sections of parasitized erythrocytes (PE) by electron microscopy. We find that actin dynamics play multiple, important roles in the hemoglobin transport pathway, and that hemoglobin delivery to the FV via the cytostomes might be required for parasite survival. Evidence is provided for a new model, in which hemoglobin transport to the FV occurs by a vesicle-independent process.

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“…However, cytoskeletal materials and peripheral proteins may remain on the cytoplasmic surface with such treatment (Shurety et al 1998;Avery et al 2000;Wetzel et al 2003). Such residue is thought to obstruct observation of the membrane proteins.…”

Section: Membrane Preparation Processmentioning

confidence: 99%

Imaging by Atomic Force Microscopy of the Plasma Membrane of Prestin-Transfected Chinese Hamster Ovary Cells

Murakoshi

Gomi

Iida

et al. 2006

JARO

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The high sensitivity of mammalian hearing is achieved by amplification of the motion of the cochlear partition. This cochlear amplification is thought to be generated by the elongation and contraction of outer hair cells (OHCs) in response to acoustical stimulation. This motility is made possible by a membrane protein embedded in the lateral membrane of OHCs. Although a fructose transporter, GLUT-5, was initially proposed to be this protein, a later study identified the gene of the motor protein distributed throughout the OHC plasma membrane. This protein has been named Bprestin.^However, although previous morphological studies by electron microscopy and atomic force microscopy (AFM) found the lateral wall of OHCs to be covered with 10-nm particles, believed to be motor proteins, it is unknown whether such particles consist only of prestin or are a complex of GLUT-5 and prestin molecules. To determine if the 10-nm particles are indeed constituted only of prestin, plasma membranes of prestin-transfected and untransfected Chinese hamster ovary (CHO) cells, which do not express GLUT-5, were observed by AFM. First, the cells attached to a substrate were sonicated so that only the plasma membrane remained on the substrate. The cytoplasmic face of the cell was observed by the tapping mode of the AFM in liquid. As a result, particle-like structures were recognized on the plasma membranes of both the prestintransfected and untransfected CHO cells. Comparison of the difference in the frequency distribution of these structures between those two cells showed approximately 75% of the particle-like structures with a diameter of 8-12 nm in the prestin-transfected CHO cells to be possibly constituted only by prestin molecules. Our data suggest that the densely packed 10-nm particles observed on the OHC lateral wall are likely to be constituted only of prestin molecules.

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Fluid-Phase Markers in the Basolateral Endocytic Pathway Accumulate in Response to the Actin Assembly-promoting Drug Jasplakinolide

Cited by 100 publications

References 51 publications

Effects of Jasplakinolide on the Kinetics of Actin Polymerization

Effects of Jasplakinolide on the Kinetics of Actin Polymerization

A new model for hemoglobin ingestion and transport by the human malaria parasite Plasmodium falciparum

Imaging by Atomic Force Microscopy of the Plasma Membrane of Prestin-Transfected Chinese Hamster Ovary Cells

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