Calmodulin Bound to the First IQ Motif Is Responsible for Calcium-dependent Regulation of Myosin 5a

Lu, Zhenwu; Shen, Mei; Cao, Yang; Zhang, Hai-Man; Yao, Linlin; Li, Xiangdong

doi:10.1074/jbc.m112.343079

Cited by 28 publications

(44 citation statements)

References 37 publications

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“…We showed previously that Myo5a-GTD inhibits not only the ATPase activity of Myo5a-HMM but also that of Myo5a-S1 containing the motor domain and the IQ1 motif (34). Similarly, we found that Myo5c-GTD inhibits the ATPase activity of Myo5c-S1 in a Ca 2ϩ -dependent manner (Fig.…”

Section: The Atpase Activity Of Myo5c Is Inhibited By Its Gtd In a Casupporting

confidence: 73%

“…Thus, CaM, particularly the C-lobe, should have a different conformation when bound to the IQ1 of Myo5a or to the IQ1 of Myo5c. Previously, our biochemical analyses have shown that the C-lobe of CaM bound to IQ1 of Myo5a is critical for the interaction with the GTD (34). We therefore proposed that the H11-H12 loop of the GTD interacts with the C-lobe of CaM bound to the IQ1 motif (Fig.…”

Section: Discussionmentioning

confidence: 88%

“…Expression and Purification of Myo5-HMM and -S1-The cDNAs of mouse Myo5a-HMM (amino acids 1-1234) and -S1 (amino acids 1-791) were subcloned into pFastHFTc (a modified pFastHTc vector encoding an N-terminal His tag and FLAG tag) as described previously (34). The cDNAs of human Myo5c-HMM (amino acids 1-1129) and -S1 (amino acids 1-781) were subcloned into pFastHFTb as described previously (31).…”

Section: Methodsmentioning

confidence: 99%

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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

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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.

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Section: The Atpase Activity Of Myo5c Is Inhibited By Its Gtd In a Casupporting

confidence: 73%

Section: Discussionmentioning

confidence: 88%

Section: Methodsmentioning

confidence: 99%

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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

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“…Oligonucleotides were synthesized by Invitrogen. Actin, CaM, and MLCK (myosin light chain kinase) were prepared as described previously (22,23). Two mouse myosin-5a constructs, i.e.…”

Section: Methodsmentioning

confidence: 99%

“…Two mouse myosin-5a constructs, i.e. Myo5a-1IQ (containing the motor domain and 1st IQ, amino acids 1-791) and Myo5a-⌬T (myosin-5a with the globular tail domain deleted), were prepared as described (23,24). SkM (skeletal muscle myosin) was prepared from rabbit muscle according to a published method (25).…”

Section: Methodsmentioning

confidence: 99%

Mouse Myosin-19 Is a Plus-end-directed, High-duty Ratio Molecular Motor

Zhang

et al. 2014

Journal of Biological Chemistry

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Background: Myosin-19 is strongly associated with mitochondria and plays a role in the transport of mitochondria. Results: The light chains of myosin-19 are the regulatory light chains of myosin-2. ADP release is rate-limiting for acto-Myo19 ATPase and ADP strongly inhibits myosin-19 motor function. Conclusion: Myosin-19 is a plus-end-directed, high-duty ratio molecular motor. Significance: Myosin-19 functions as a molecular motor.

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Creation of a myosin Va‐TAP‐tagged mouse and identification of potential myosin Va‐interacting proteins in the cerebellum

et al. 2018

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The actin-based motor myosin Va transports numerous cargos, including the smooth endoplasmic reticulum (SER) in cerebellar Purkinje neurons (PNs) and melanosomes in melanocytes. Identifying proteins that interact with this myosin is key to understanding its cellular functions. Toward that end, we used recombineering to insert via homologous recombination a tandem affinity purification (TAP) tag composed of the immunoglobulin G-binding domain of protein A, a tobacco etch virus cleavage site, and a FLAG tag into the mouse MYO5A locus immediately after the initiation codon. Importantly, we provide evidence that the TAP-tagged version of myosin Va (TAP-MyoVa) functions normally in terms of SER transport in PNs and melanosome positioning in melanocytes. Given this and other evidence that TAP-MyoVa is fully functional, we purified it together with associated proteins directly from juvenile mouse cerebella and subjected the samples to mass spectroscopic analyses. As expected, known myosin Va-binding partners like dynein light chain were identified. Importantly, numerous novel interacting proteins were also tentatively identified, including guanine nucleotide-binding protein G(o) subunit alpha (Gnao1), a biomarker for schizophrenia. Consistently, an antibody to Gnao1 immunoprecipitates myosin Va, and Gnao1's localization to PN dendritic spines depends on myosin Va. The mouse model created here should facilitate the identification of novel myosin Va-binding partners, which in turn should advance our understanding of the roles played by this important myosin in vivo.

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Calmodulin Bound to the First IQ Motif Is Responsible for Calcium-dependent Regulation of Myosin 5a

Cited by 28 publications

References 37 publications

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

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

Mouse Myosin-19 Is a Plus-end-directed, High-duty Ratio Molecular Motor

Creation of a myosin Va‐TAP‐tagged mouse and identification of potential myosin Va‐interacting proteins in the cerebellum

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