Monomeric myosin V uses two binding regions for the assembly of stable translocation complexes

Heuck, Alexander; Du, Tung-Gia; Jellbauer, Stephan; Richter, Klaus; Kruse, Claudia; Jaklin, Sigrun; Müller, Marisa; Büchner, Johannes; Jansen, Ralf‐Peter; Niessing, Dierk

doi:10.1073/pnas.0706780104

Cited by 29 publications

(57 citation statements)

References 40 publications

(67 reference statements)

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“…Recent publications demonstrate that She3p binds the rod region of Myo4p (Heuck et al, 2007;Hodges et al, 2008). Thus, Myo4p attaches to cargoes via a mechanism that is distinct from Myo2p and currently is not well established.…”

Section: Ptc1 Affects Both Myosin-v Motors and Organelle-specific Recmentioning

confidence: 99%

“…PTC1-NBP2 regulates Myo4p function through the CWI pathway (Du et al, 2006). Myo2p forms a dimer (Beningo et al, 2000), and Myo4p forms a single-head motor (Dunn et al, 2007;Heuck et al, 2007;Hodges et al, 2008). The location of cargo attachment to Myo4p is currently unknown.…”

Section: Acknowledgmentsmentioning

confidence: 99%

“…Similarly, the cortical ER is moved by Myo4p, the other yeast myosin-V, which forms a complex with She3p (Estrada et al, 2003). Several mRNAs are also moved by Myo4p and require both She3p and She2p, proteins whose roles are not well established (Bohl et al, 2000;Long et al, 2000;Shepard et al, 2003;Heuck et al, 2007;Hodges et al, 2008). Moreover, rather than attachment to the globular tail, She3p binds the rod of Myo4p.…”

Section: Introductionmentioning

confidence: 99%

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PTC1Is Required for Vacuole Inheritance and Promotes the Association of the Myosin-V Vacuole-specific Receptor Complex

Jin

Eves

Tang

et al. 2009

MBoC

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Organelle inheritance occurs during cell division. In Saccharomyces cerevisiae, inheritance of the vacuole, and the distribution of mitochondria and cortical endoplasmic reticulum are regulated by Ptc1p, a type 2C protein phosphatase. Here we show that PTC1/VAC10 controls the distribution of additional cargoes moved by a myosin-V motor. These include peroxisomes, secretory vesicles, cargoes of Myo2p, and ASH1 mRNA, a cargo of Myo4p. We find that Ptc1p is required for the proper distribution of both Myo2p and Myo4p. Surprisingly, PTC1 is also required to maintain the steady-state levels of organelle-specific receptors, including Vac17p, Inp2p, and Mmr1p, which attach Myo2p to the vacuole, peroxisomes, and mitochondria, respectively. Furthermore, Vac17p fused to the cargo-binding domain of Myo2p suppressed the vacuole inheritance defect in ptc1⌬ cells. These findings suggest that PTC1 promotes the association of myosin-V with its organelle-specific adaptor proteins. Moreover, these observations suggest that despite the existence of organelle-specific receptors, there is a higher order regulation that coordinates the movement of diverse cellular components. INTRODUCTIONDuring each cell cycle, cytoplasmic organelles are actively distributed between dividing cells to maintain organelle copy number and volume. The yeast Saccharomyces cerevisiae is an excellent model system for studying the spatial and temporal control of organelle inheritance. In yeast, several organelles are transmitted from mother to daughter cells. These include the vacuole, mitochondria, the endoplasmic reticulum (ER), late-Golgi elements and peroxisomes (reviewed in Weisman, 2006).Early in the cell cycle, a portion of each organelle is transported into the emerging bud. The polarized transport of most organelles from the mother to the bud is an active process that requires the actin cytoskeleton, myosin-V motors, and receptor proteins, which physically connect the motor to organelle cargoes (Beach et al., 2000;Yin et al., 2000;Boldogh et al., 2001;Barr, 2002;Bretscher, 2003;Du et al., 2004;Fagarasanu et al., 2006bFagarasanu et al., , 2007Weisman, 2006). Thus, formation of a complex between the motor and receptor protein is important for polarized organelle transport.Similar to yeast, in vertebrates, myosin-V motors move cargoes along the actin cytoskeleton. The best studied cargo of vertebrate myosin-V are melanosomes, which are moved in melanocytes by myosin-Va. Melanosomes attach to myosin-Va through Rab27a and melanophilin (Fukuda and Kuroda, 2002;Wu et al., 2002). Rab27a, through geranylgeranylation, attaches to the melanosome membrane, and melanophilin connects myosin-Va and Rab27a. Myosin-V-based intracellular movement has been analyzed in many other eukaryotes, including frogs, fish, mammals, and plants; plant myosin-XI is the functional homologue of yeast and vertebrate myosin-V ( Kinkema and Schiefelbein, 1994;Provance and Mercer, 1999;Tuma and Gelfand, 1999;Desnos et al., 2007;Li and Nebenfuhr, 2007;Sheets et al., 2007;Shimmen, 2007).Ge...

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Section: Ptc1 Affects Both Myosin-v Motors and Organelle-specific Recmentioning

confidence: 99%

Section: Acknowledgmentsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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PTC1Is Required for Vacuole Inheritance and Promotes the Association of the Myosin-V Vacuole-specific Receptor Complex

Jin

Eves

Tang

et al. 2009

MBoC

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“…Once in the cytoplasm, the ASH1 precomplex interacts with another subcomplex consisting of the type V myosin motor myosin 4 (Myo4p) and the RNA-binding protein She3p. The nuclear and cytoplasmic precomplexes form the mature transport complex via the direct interaction between She2p and She3p and their synergistic binding to the zip-code elements of the ASH1 mRNA (8,(23)(24)(25)(26)(27)(28).…”

Section: Significancementioning

confidence: 99%

Role of Loc1p in assembly and reorganization of nuclear ASH1 messenger ribonucleoprotein particles in yeast

Niedner

Müller

Moorthy

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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Directional transport of mRNA is a universal feature in eukaryotes, requiring the assembly of motor-dependent RNA-transport particles. The cytoplasmic transport of mRNAs is preceded by the nuclear assembly of pre-messenger ribonucleoprotein particles (mRNPs). In budding yeast, the asymmetric synthesis of HO 1 (ASH1) pre-mRNP originates already cotranscriptionally and passes through the nucleolus before its nuclear export. The nucleolar localization of ASH1 mRNA protein 1 (Loc1p) is required for efficient ASH1 mRNA localization. Immunoprecipitation experiments have revealed that Loc1p forms cocomplexes with other components of the ASH1 transport complex. However, it remains unclear how Loc1p is recruited into this mRNP and why Loc1p is important for ASH1 mRNA localization. Here we demonstrate that Loc1p undergoes a direct and specific interaction with the ASH1 mRNA-binding Swi5p-dependent HO expression protein 2 (She2p). This cocomplex shows higher affinity and specificity for RNA bearing localization elements than the individual proteins. It also stabilizes the otherwise transient binding of She2p to ASH1 mRNA, suggesting that cooperative mRNA binding of Loc1p with She2p is the required nuclear function of Loc1p for ASH1 mRNA localization. After nuclear export, myosin-bound She3p joins the ASH1 mRNP to form a highly specific cocomplex with She2p and ASH1 mRNA. Because Loc1p is found only in the nucleus, it must be removed from the complex directly before or after export. In vitro and in vivo experiments indicate that the synergistic interaction of She2p and She3p displaces Loc1p from the ASH1 complex, allowing free Loc1p to rapidly reenter the nucle(ol)us. Together these findings suggest an ordered process of nuclear assembly and reorganization for the maturation of localizing ASH1 mRNPs.M essenger RNA localization is a universal feature of eukaryotes (1-3). By complementing transcriptional control (4), it fulfills a variety of functions, including the establishment of cell polarity and specialization of subcellular regions. In recent years, the directional transport of asymmetric synthesis of HO 1 (ASH1) mRNA in budding yeast has emerged as a particularly well-suited model to study mechanistic principles of RNA localization. Here, comparably few proteins participate in the directional transport of ASH1 mRNA and about 30 other transcripts (5, 6).Chromatin-immunoprecipitaton experiments revealed that the dedicated RNA-binding Swi5p-dependent HO expression protein 2 (She2p) binds already cotranscriptionally to nascent ASH1 mRNA (7, 8). Two additional RNA-binding proteins, pumiliohomology domain family protein 6 (Puf6p) and heterogeneous nuclear RNP K-like protein 1 (Khd1p), are also present in the nucleus, bind to ASH1 mRNA, and act in the cytoplasm as translational repressors during ASH1 transport (9-12). A fourth nuclear factor, termed localization of ASH1 mRNA protein 1 (Loc1p), has been implicated in the assembly of nuclear premessenger ribonucleoprotein particles (mRNPs). Like Puf6p, Loc1p is a nuclear protei...

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“…crystal structure | LPGV hook | type II turn | motor assembly M yosin 4 protein (Myo4p) of Saccharomyces cerevisiae is a monomeric myosin unable to move processively on actin filaments unless dimerized (1)(2)(3). Dimerization is regulated by the availability of cargo (4)(5)(6).…”

mentioning

confidence: 99%

Hooking She3p onto She2p for myosin-mediated cytoplasmic mRNA transport

Singh

Blobel

Shi

2014

Proc. Natl. Acad. Sci. U.S.A.

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The segregation of approximately two dozen distinct mRNAs from yeast mother to daughter cell cytoplasm is a classical paradigm for eukaryotic mRNA transport. The information for transport resides in an mRNA element 40–100 nt in length, known as “zipcode.” Targeted transport requires properly positioned actin filaments and cooperative loading of mRNA cargo to myosin. Cargo loading to myosin uses myosin 4 protein (Myo4p), swi5p-dependent HO expression 2 protein (She2p) and 3 protein (She3p), and zipcode. We previously determined a crystal structure of Myo4p and She3p, their 1:2 stoichiometry and interactome; we furthermore showed that the motor complex assembly requires two Myo4p⋅She3p heterotrimers, one She2p tetramer, and at least a single zipcode to yield a stable complex of [Myo4p⋅She3p⋅She2p⋅zipcode] in 2:4:4:1 stoichiometry in vitro. Here, we report a structure at 2.8-Å resolution of a cocrystal of a She2p tetramer bound to a segment of She3p. In this crystal structure, the She3p segment forms a striking hook that binds to a shallow hydrophobic pocket on the surface of each She2p subunit of the tetramer. Both She3p hook and cognate She2p binding pocket are composed of highly conserved residues. We also discovered a highly conserved region of She3p upstream of its hook region. Because this region consists of basic and aromatic residues, it likely represents part of She3p’s binding activity for zipcode. Because She2p also exhibits zipcode-binding activity, we suggest that “hooking” She3p onto She2p aligns each of their zipcode-binding activities into a high-affinity site, thereby linking motor assembly to zipcode.

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Monomeric myosin V uses two binding regions for the assembly of stable translocation complexes

Cited by 29 publications

References 40 publications

PTC1Is Required for Vacuole Inheritance and Promotes the Association of the Myosin-V Vacuole-specific Receptor Complex

PTC1Is Required for Vacuole Inheritance and Promotes the Association of the Myosin-V Vacuole-specific Receptor Complex

Role of Loc1p in assembly and reorganization of nuclear ASH1 messenger ribonucleoprotein particles in yeast

Hooking She3p onto She2p for myosin-mediated cytoplasmic mRNA transport

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