Myosin Va Bound to Phagosomes Binds to F-Actin and Delays Microtubule-dependent Motility

Al-Haddad, Ahmed; Shonn, Marion A.; Redlich, Bärbel; Blocker, Ariel; Burkhardt, Janis K.; Yu, Hanry; Hammer, John A.; Weiss, Dieter G.; Walter, Steffen; Griffiths, Gareth; Kuznetsov, Sergei A.

doi:10.1091/mbc.12.9.2742

Cited by 93 publications

(91 citation statements)

References 56 publications

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“…8). Evidence for a dual transport system has also been found for melanosomes (Brown, 1999;Reck-Peterson et al, 2000), the endoplasmic reticulum (Tabb et al, 1998;Reck-Peterson et al, 2000) and phagosomes (Al-Haddad et al, 2001). Along these lines, it appears that motor proteins associated with the surface of organelles, are often organised in protein complexes (Schliwa, 1999).…”

Section: Existence and Recruitment Of Motor Protein Complexesmentioning

confidence: 94%

Myosin Va facilitates the distribution of secretory granules in the F-actin rich cortex of PC12 cells

Rudolf

Kögel

Kuznetsov

et al. 2003

Journal of Cell Science

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Neuroendocrine secretory granules, the storage organelles for neuropeptides and hormones, are formed at the transGolgi network, stored inside the cell and exocytosed upon stimulation. Previously, we have reported that newly formed secretory granules of PC12 cells are transported in a microtubule-dependent manner from the trans-Golgi network to the F-actin-rich cell cortex, where they undergo short directed movements and exhibit a homogeneous distribution. Here we provide morphological and biochemical evidence that myosin Va is associated with secretory granules. Expression of a dominant-negative tail domain of myosin Va in PC12 cells led to an extensive clustering of secretory granules close to the cell periphery, a loss of their cortical restriction and a strong reduction in their motility in the actin cortex. Based on this data we propose a model that implies a dual transport system for secretory granules: after microtubule-dependent delivery to the cell periphery, secretory granules exhibit a myosin Va-dependent transport leading to their restriction and even dispersal in the F-actin-rich cortex of PC12 cells. Movie available online

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Section: Existence and Recruitment Of Motor Protein Complexesmentioning

confidence: 94%

Myosin Va facilitates the distribution of secretory granules in the F-actin rich cortex of PC12 cells

Rudolf

Kögel

Kuznetsov

et al. 2003

Journal of Cell Science

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120

131

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“…The primary P2X7R-knockout and WT bone marrow macrophages were isolated as described (Al-Haddad et al, 2001) from wild-type and P2X7R -/-C57 BL/6 mice (Solle et al, 2001). …”

Section: Methodsmentioning

confidence: 99%

Lipids regulate P2X7-receptor-dependent actin assembly by phagosomes via ADP translocation and ATP synthesis in the phagosome lumen

Rybin

Anand

Anes

et al. 2009

Journal of Cell Science

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Latex bead phagosomes isolated from J774 macrophages polymerize actin. We show here that five lipids – phosphatidylinositol-4-phosphate, phosphatidylinositol-(4,5)-bisphosphate, sphingosine-1-phosphate (S1P), ceramide-1-phosphate and phosphatidic acid – stimulate both actin assembly and transport of ADP across the phagosomal membrane into the lumen. Once there, this ADP is converted to ATP by adenylate kinase activity. High luminal ATP concentrations correlated well with phagosome actin assembly under different conditions. The ATP-binding P2X7 receptor (P2X7R) was detected in phagosomes. Although S1P stimulated actin assembly by phagosomes from P2X7R-containing bone marrow macrophages, S1P-stimulated actin assembly was inhibited in phagosomes from cells lacking P2X7R. We propose that luminal ATP accumulates in response to selected lipids and activates the P2X7R that signals across the phagosomal membrane to trigger actin assembly on the cytoplasmic membrane surface. In the accompanying paper by Kuehnel et al. (doi:10.1242/jcs.034207), more evidence is provided in support of this model from the analysis of actin assembly at the plasma membrane of intact macrophages.

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“…The primary neuronal defect thought to be responsible is the absence of smooth endoplasmic reticulum (SER) within the dendritic spines of Purkinje neurons (Takagishi et al, 1996). Analyses of organelle movement in cultured melanocytes (Provance et al, 1996;Nascimento et al, 1997;Wu et al, 1997Wu et al, , 1998a, neurons (Bridgman, 1999), and macrophages (Al-Haddad et al, 2001) from the dilute and wild-type mice have provided evidence for a direct role for Myo5a in both organelle transport and tethering. Moreover, in all three cell types, normal organelle movement requires participation of MT based motors as well as Myo5a.…”

Section: Introductionmentioning

confidence: 99%

Myosin-Va Binds to and Mechanochemically Couples Microtubules to Actin Filaments

Cao

Chang

Masters

et al. 2004

MBoC

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Myosin-Va was identified as a microtubule binding protein by cosedimentation analysis in the presence of microtubules. Native myosin-Va purified from chick brain, as well as the expressed globular tail domain of this myosin, but not head domain bound to microtubule-associated protein-free microtubules. Binding of myosin-Va to microtubules was saturable and of moderately high affinity (ϳ1:24 Myosin-Va:tubulin; K d ‫؍‬ 70 nM). Myosin-Va may bind to microtubules via its tail domain because microtubule-bound myosin-Va retained the ability to bind actin filaments resulting in the formation of cross-linked gels of microtubules and actin, as assessed by fluorescence and electron microscopy. In low Ca 2؉ , ATP addition induced dissolution of these gels, but not release of myosin-Va from MTs. However, in 10 M Ca 2؉ , ATP addition resulted in the contraction of the gels into aster-like arrays. These results demonstrate that myosin-Va is a microtubule binding protein that cross-links and mechanochemically couples microtubules to actin filaments. INTRODUCTIONMyosin-Va (Myo5a) is the best characterized of three class V myosins identified in vertebrates (Berg et al., 2001). Myo5a is a two-headed motor, consisting of two heavy chains and multiple light chains (reviewed in Reck-Peterson et al., 2000). The N-terminal half of each heavy chain consists of a head or motor domain followed by a neck domain composed of six IQ motifs, each of which binds a calmodulin light chain although chicken Myo5a also contains a pair of myosin-II essential light chains . The C-terminal tail domain consists of a proximal dimerization stalk of largely coiled-coil forming alpha helix followed by a globular domain that has been shown to interact with numerous presumed "cargo" molecules (Reck-Peterson et al., 2000;Karcher et al., 2002;Langford, 2002). The tail domain also contains at least a pair of the low-molecular-weight light chain, LC8 or PIN, first identified as a light chain of the microtuble (MT) motor, dynein, but subsequently shown to interact with numerous other proteins . Biochemical studies on both tissue-purified and baculovirus-expressed Myo5a (reviewed in Reck-Peterson et al., 2000;Mehta, 2001) have shown that it is a Ca 2ϩ -regulated, barbed-end directed, processive motor that takes large, ϳ36-nm steps in a hand-overhand manner (Forkey et al., 2003; Yildiz et al., 2003) along the actin filament.Insights into the potential functions for Myo5a have come from several lines of investigation, including identification of organelles and proteins that interact with Myo5a (ReckPeterson et al., 2000;Karcher et al., 2002;Langford, 2002). The most powerful strategy has been the phenotypic characterization of lethal alleles of the dilute (Myo5a lethal mutant) mouse and cells derived from them. The dilute mice develop severe seizures and die within 3 wk after birth. The primary neuronal defect thought to be responsible is the absence of smooth endoplasmic reticulum (SER) within the dendritic spines of Purkinje neurons (Takagishi et al., 1996). Anal...

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Myosin Va Bound to Phagosomes Binds to F-Actin and Delays Microtubule-dependent Motility

Cited by 93 publications

References 56 publications

Myosin Va facilitates the distribution of secretory granules in the F-actin rich cortex of PC12 cells

Myosin Va facilitates the distribution of secretory granules in the F-actin rich cortex of PC12 cells

Lipids regulate P2X7-receptor-dependent actin assembly by phagosomes via ADP translocation and ATP synthesis in the phagosome lumen

Myosin-Va Binds to and Mechanochemically Couples Microtubules to Actin Filaments

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