Myosin VI Contains a Compact Structural Motif that Binds to Ubiquitin Chains

He, Fahu; Wollscheid, Hans-Peter; Nowicka, Urszula; Biancospino, Matteo; Valentini, Eleonora; Ehlinger, Aaron; Acconcia, Filippo; Magistrati, Elisa; Polo, Simona; Walters, Kylie J.

doi:10.1016/j.celrep.2016.01.079

Cited by 50 publications

(68 citation statements)

References 52 publications

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“…Lack of activation is consistent with recent studies showing that the large insert in the myosin VI tail domain, which is present in our construct, occludes the OPTN binding site (49). Furthermore, OPTN must be ubiquitinated before binding myosin VI (50).…”

Section: Lovdab Controls Myosin VI Recruitment With High Spatial and supporting

confidence: 92%

Investigations of human myosin VI targeting using optogenetically controlled cargo loading

French

Sosnick

Rock

2017

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

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Myosins play countless critical roles in the cell, each requiring it to be activated at a specific location and time. To control myosin VI with this specificity, we created an optogenetic tool for activating myosin VI by fusing the light-sensitive Avena sativa phototropin1 LOV2 domain to a peptide from Dab2 (LOVDab), a myosin VI cargo protein. Our approach harnesses the native targeting and activation mechanism of myosin VI, allowing direct inferences on myosin VI function. LOVDab robustly recruits human full-length myosin VI to various organelles in vivo and hinders peroxisome motion in a lightcontrollable manner. LOVDab also activates myosin VI in an in vitro gliding filament assay. Our data suggest that protein and lipid cargoes cooperate to activate myosin VI, allowing myosin VI to integrate Ca 2+, lipid, and protein cargo signals in the cell to deploy in a site-specific manner.otor proteins play countless roles in biology, each requiring the motor to be recruited and activated at a particular time and place inside the cell. To dissect these multiple roles, we must develop tools that allow us to control the recruitment and activation process. One promising technique for achieving this goal is through optogenetics (1). Optogenetics involves the engineering and application of optically controlled, genetically encoded proteins and is transforming the fields of neuro-and cell biology (2, 3). A major benefit of optogenetics is that proteins are activated using light, which allows for high temporal and spatial control over a protein of interest.Myosin VI is a motor protein whose study could particularly benefit from optogenetic control. It is the only myosin known to walk toward the pointed end of actin filaments (4, 5). This property enables it to perform a diverse array of cellular functions, including cell division, endosome trafficking, autophagy, and Golgi and plasma membrane anchoring (6-9). Myosin VI is also autoinhibited, a property that is commonly found in other myosins (10). When myosin VI binds to cargo through specialized adaptor proteins, this autoinhibition is relieved through a poorly understood mechanism likely involving the disruption of an interaction between its cargo binding domain (CBD) and the myosin head (11,12). Dissociation of the CBD from the head both frees the head to bind tightly to actin and exposes dimerization sites throughout the tail domain of myosin VI, allowing it to become a processive dimer (13-15). In some cases, myosin VI could conceivably function as a monomer, for example when fulfilling its role as a membrane tether during spermatid individualization (16). If this is the case, more work is needed to elucidate the cellular signals that determine its oligomeric state at each site of action.Myosin VI has two classes of cargo proteins that bind to distinct, conserved motifs on the myosin VI C terminus (17). Disabled2, or Dab2, belongs to the class of cargo proteins that bind to a conserved WWY site on myosin (18). Optineurin (OPTN) is a member of the second class that binds ...

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Section: Lovdab Controls Myosin VI Recruitment With High Spatial and supporting

confidence: 92%

Investigations of human myosin VI targeting using optogenetically controlled cargo loading

French

Sosnick

Rock

2017

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

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“…Ubiquitination generally regulates cellular function by timely and selective addition of the ubiquitin molecule to its substrate (33). There are eight different types of The first seven types include K6, K11, K27, K29, K33, K48, and K63, which are associated with the internal lysine K and the glycine G at the C terminus of ubiquitin molecules (34)(35)(36). Most of the studies have investigated K48 and K63 polyadenylation modification, in which the polyubiquitination modification at position K48 mainly plays a role in degradation (37,38) and the polyubiquitination modification at position K63 mainly plays a role in signal transduction DNA repair function (39,40).…”

Section: Discussionmentioning

confidence: 99%

The Superimposed Deubiquitination Effect of OTULIN and Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Nsp11 Promotes Multiplication of PRRSV

Shi

Zhang

et al. 2018

J Virol

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Linear ubiquitination plays an important role in the regulation of the immune response by regulating nuclear factor κB (NF-κB). The linear ubiquitination-specific deubiquitinase ovarian tumor domain deubiquitinase with linear linkage specificity (OTULIN) can control the immune signaling transduction pathway by restricting the Met1-linked ubiquitination process. In our study, the porcine OTLLIN gene was cloned and deubiquitin functions were detected in a porcine reproductive and respiratory syndrome virus (PRRSV)-infected-cell model. PRRSV infection promotes the expression of the OTULIN gene; in turn, overexpression of OTULIN contributes to PRRSV proliferation. There is negative regulation of innate immunity with OTULIN during viral infection. The cooperative effects of swine OTULIN and PRRSV Nsp11 potentiate the ability to reduce levels of cellular protein ubiquitin associated with innate immunity. Importantly, PRRSV Nsp11 recruits OTULIN through a nonenzymatic combination to enhance its ability to remove linear ubiquitination targeting NEMO, resulting in a superimposed effect that inhibits the production of type I interferons (IFNs). Our report presents a new model of virus utilization of the ubiquitin-protease system from the perspective of the viral proteins that interact with cell deubiquitination enzymes, providing new ideas for prevention and control of PRRSV. Deubiquitination effects of swine OTULIN were identified. The interaction between porcine OTULIN and PRRSV Nsp11 is dependent on the OTU domain. PRRSV Nsp11 recruits OTULIN through a nonenzymatic combination to promote removal of linear ubiquitination targeting NEMO, resulting in a superimposed effect that inhibits the production of type I IFNs.

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“…Notably, the large Phe4 CSP for K63 proximal Ub is the only instance when we observe a significant shift in this residue. This Phe4-patch surface is also used by K63 diUb to bind the myosin VI MyUb domain (51); the surface formed by Phe4, Phe45, Ala46, Lys48 and Thr66 in proximal ubiquitin contacts the MyUb helix 1. The observation is also consistent with the important role of the Ub residue Phe4 in endocytosis (52), which was shown to be mediated by binding to K63-linked polyUbs (53).…”

Section: Discussionmentioning

confidence: 99%

Novel TDP2-ubiquitin interactions and their importance for the repair of topoisomerase II-mediated DNA damage

et al. 2016

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Tyrosyl DNA phosphodiesterase 2 (TDP2) is a multifunctional protein implicated in DNA repair, signal transduction and transcriptional regulation. In its DNA repair role, TDP2 safeguards genome integrity by hydrolyzing 5′-tyrosyl DNA adducts formed by abortive topoisomerase II (Top2) cleavage complexes to allow error-free repair of DNA double-strand breaks, thereby conferring cellular resistance against Top2 poisons. TDP2 consists of a C-terminal catalytic domain responsible for its phosphodiesterase activity, and a functionally uncharacterized N-terminal region. Here, we demonstrate that this N-terminal region contains a ubiquitin (Ub)-associated (UBA) domain capable of binding multiple forms of Ub with distinct modes of interactions and preference for either K48- or K63-linked polyUbs over monoUb. The structure of TDP2 UBA bound to monoUb shows a canonical mode of UBA-Ub interaction. However, the absence of the highly conserved MGF motif and the presence of a fourth α-helix make TDP2 UBA distinct from other known UBAs. Mutations in the TDP2 UBA-Ub binding interface do not affect nuclear import of TDP2, but severely compromise its ability to repair Top2-mediated DNA damage, thus establishing the importance of the TDP2 UBA–Ub interaction in DNA repair. The differential binding to multiple Ub forms could be important for responding to DNA damage signals under different contexts or to support the multi-functionality of TDP2.

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Myosin VI Contains a Compact Structural Motif that Binds to Ubiquitin Chains

Cited by 50 publications

References 52 publications

Investigations of human myosin VI targeting using optogenetically controlled cargo loading

Investigations of human myosin VI targeting using optogenetically controlled cargo loading

The Superimposed Deubiquitination Effect of OTULIN and Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Nsp11 Promotes Multiplication of PRRSV

Novel TDP2-ubiquitin interactions and their importance for the repair of topoisomerase II-mediated DNA damage

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