Structural Insights into Functional Overlapping and Differentiation among Myosin V Motors

Nascimento, Andrey Fabricio Ziem; Trindade, Daniel Maragno; Tonoli, C.C.C.; Giuseppe, P.O.; Assis, Leandro H. P.; Honorato, Rodrigo Vargas; Oliveira, Paulo Sérgio Lopes de; Mahajan, P.; Burgess-Brown, N.; Delft, Frank von; Larson, Roy Edward; Murakami, Mário Tyago

doi:10.1074/jbc.m113.507202

Cited by 30 publications

(64 citation statements)

References 68 publications

(68 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Several Myo-V isoforms has been reported (Nascimento et al, 2013) and it has been established that the Myo-Vb is implicated in the basolateral trafficking of voltage-gated K + channel Kv1.5 (Schumacher-Bass et al, 2014). Interestingly, Myo-Vc is highly expressed in epithelial (Rodriguez and Cheney, 2002; Jacobs et al, 2009; Nascimento et al, 2013), and this motor protein is known to interact with Rab8 (Jacobs et al, 2009; Xu et al, 2009) which has been reported to play a role in the trafficking of KCa3.1 in epithelial cells (Bertuccio et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

The Role of the Cytoskeleton and Myosin-Vc in the Targeting of KCa3.1 to the Basolateral Membrane of Polarized Epithelial Cells

Farquhar¹,

Rodrigues²,

Hamilton³

2017

Front. Physiol.

View full text Add to dashboard Cite

Understanding the targeting of KCa3.1 to the basolateral membrane (BLM) of polarized epithelial cells is still emerging. Here, we examined the role of the cytoskeleton (microtubules and microfilaments) and Myosin-Vc (Myo-Vc) in the targeting of KCa3.1 in Fischer rat thyroid epithelial cells. We used a pharmacological approach with immunoblot (for the BLM expression of KCa3.1), Ussing chamber (functional BLM expression of KCa3.1) and siRNA experiments. The actin cytoskeleton inhibitors cytochalasin D (10 μM, 5 h) and latrunculin A (10 μM, 5 h) reduced the targeting of KCa3.1 to the BLM by 88 ± 4 and 70 ± 5%, respectively. Colchicine (10 μM, 5 h) a microtubule inhibitor reduced targeting of KCa3.1 to the BLM by 63 ± 7% and decreased 1-EBIO-stimulated KCa3.1 K+ current by 46 ± 18%, compared with control cells. ML9 (10 μM, 5 h), an inhibitor of myosin light chain kinase, decreased targeting of the channel by 83 ± 2% and reduced K+ current by 54 ± 8% compared to control cells. Inhibiting Myo-V with 2,3-butanedione monoxime (10 mM, 5 h) reduced targeting of the channel to the BLM by 58 ± 5% and decreased the stimulated current of KCa3.1 by 48 ± 12% compared with control cells. Finally, using siRNA for Myo-Vc, we demonstrated that knockdown of Myo-Vc reduced the BLM expression of KCa3.1 by 44 ± 7% and KCa3.1 K+ current by 1.04 ± 0.14 μA compared with control cells. These data suggest that the microtubule and microfilament cytoskeleton and Myo-Vc are critical for the targeting of KCa3.1.

show abstract

Section: Discussionmentioning

confidence: 99%

The Role of the Cytoskeleton and Myosin-Vc in the Targeting of KCa3.1 to the Basolateral Membrane of Polarized Epithelial Cells

Farquhar¹,

Rodrigues²,

Hamilton³

2017

Front. Physiol.

View full text Add to dashboard Cite

show abstract

“…We previously proposed a model of GTD-head interaction by manually docking yeast Myo2p-GTD onto the head of Myo5a. Recently, a similar model was independently proposed by Velvarska and Niessing (17) and by Nascimento et al (16). Here we present the model of GTD-head interaction (Fig.…”

Section: Discussionmentioning

confidence: 93%

“…Myo2p and Myo4p) and three types of vertebrate Myo5 (i.e. Myo5a, 5b, and 5c), have been solved (12)(13)(14)(15)(16)(17). The structure of GTD is mainly composed of ␣ helices connected by short and long loops (Fig.…”

mentioning

confidence: 99%

Identification of the Isoform-specific Interactions between the Tail and the Head of Class V Myosin

Yao

Shen

et al. 2016

Journal of Biological Chemistry

View full text Add to dashboard Cite

Vertebrates have three isoforms of class V myosin (Myo5), Myo5a, Myo5b, and Myo5c, which are involved in transport of multiple cargoes. It is well established that the motor functions of Myo5a and Myo5b are regulated by a tail inhibition mechanism. Here we found that the motor function of Myo5c was also inhibited by its globular tail domain (GTD), and this inhibition was abolished by high Ca 2؉ , indicating that the tail inhibition mechanism is conserved in vertebrate Myo5. Interestingly, we found that Myo5a-GTD and Myo5c-GTD were not interchangeable in terms of inhibition of motor function, indicating isoform-specific interactions between the GTD and the head of Myo5. To identify the isoform-specific interactions, we produced a number of Myo5 chimeras by swapping the corresponding regions of Myo5a and Myo5c. We found that Myo5a-GTD, with its H11-H12 loop being substituted with that of Myo5c, was able to inhibit the ATPase activity of Myo5c and that Myo5a-GTD was able to inhibit the ATPase activity of Myo5c-S1 and Myo5c-HMM only when their IQ1 motif was substituted with that of Myo5a. Those results indicate that the H11-H12 loop in the GTD and the IQ1 motif in the head dictate the isoformspecific interactions between the GTD and head of Myo5. Because the IQ1 motif is wrapped by calmodulin, whose conformation is influenced by the sequence of the IQ1 motif, we proposed that the calmodulin bound to the IQ1 motif interacts with the H11-H12 loop of the GTD in the inhibited state of Myo5.

show abstract

“…A binding surface on Myo4p is provided by the segment extending from 1,325-1,402, where this is structurally adjacent to the N-terminal binding surface, even though is it separated by 264 residues in the sequence. As noted before, the structure of the cargo-binding domain of Myo4p is very different from that of mammalian class V myosins where the latter exist as constitutive dimers and are regulated via a head-tail interaction (18).…”

mentioning

confidence: 99%

Structural insights into the assembly of a monomeric class V myosin

Rayment

2014

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

View full text Add to dashboard Cite

Structural Insights into Functional Overlapping and Differentiation among Myosin V Motors

Cited by 30 publications

References 68 publications

The Role of the Cytoskeleton and Myosin-Vc in the Targeting of KCa3.1 to the Basolateral Membrane of Polarized Epithelial Cells

The Role of the Cytoskeleton and Myosin-Vc in the Targeting of KCa3.1 to the Basolateral Membrane of Polarized Epithelial Cells

Identification of the Isoform-specific Interactions between the Tail and the Head of Class V Myosin

Structural insights into the assembly of a monomeric class V myosin

Contact Info

Product

Resources

About