Identification of novel MYO18A interaction partners required for myoblast adhesion and muscle integrity.
The unconventional myosin MYO18A that contains a PDZ domain is required for muscle integrity during zebrafish development. However, the mechanism by which it functions in myofibers is not clear. The presence of a PDZ domain suggests that MYO18A may interact with other partners to perform muscle-specific functions. Here we performed double-hybrid screening and co-immunoprecipitation to identify MYO18A-interacting proteins, and have identified p190RhoGEF and Golgin45 as novel partners for the MYO18A PDZ domain. We have also identified Lurap1, which was previously shown to bind MYO18A. Functional analyses indicate that, similarly as myo18a, knockdown of lurap1, p190RhoGEF and Golgin45 by morpholino oligonucleotides disrupts dystrophin localization at the sarcolemma and produces muscle lesions. Simultaneous knockdown of myo18a with either of these genes severely disrupts myofiber integrity and dystrophin localization, suggesting that they may function similarly to maintain myofiber integrity. We further show that MYO18A and its interaction partners are required for adhesion of myoblasts to extracellular matrix, and for the formation of the Golgi apparatus and organization of F-actin bundles in myoblast cells. These findings suggest that MYO18A has the potential to form a multiprotein complex that links the Golgi apparatus to F-actin, which regulates muscle integrity and function during early development.
School of Life Sciences, Shandong University, Jinan 250100, China.
Jian-Meng
JM
Cao
School of Life Sciences, Shandong University, Jinan 250100, China.
Xiao-Ning
XN
Cheng
School of Life Sciences, Shandong University, Jinan 250100, China.
Shang-Qi
SQ
Li
Department of Otolaryngology-Head &Neck Surgery; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.
Stefan
S
Heller
School of Life Sciences, Shandong University, Jinan 250100, China.
Zhi-Gang
ZG
Xu
School of Life Sciences, Shandong University, Jinan 250100, China.|Sorbonne Universités, UPMC Univ Paris 06, CNRS, Biologie du développement, Institut de Biologie Paris Seine (LBD-IBPS), 75005 Paris, France.
De-Li
DL
Shi
D000199:Actins / Q000378:metabolism; D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D000818:Animals; D002448:Cell Adhesion; D002645:Chickens; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006056:Golgi Apparatus / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D009132:Muscles / Q000502:physiology*; D032446:Myoblasts / Q000166:cytology*; D009218:Myosins / Q000737:chemistry / Q000378:metabolism*; D009841:Oligonucleotides / Q000235:genetics; D011485:Protein Binding; D000072417:Protein Domains; D025521:Tumor Suppressor Proteins / Q000378:metabolism; D020798:Two-Hybrid System Techniques; D033921:Vesicular Transport Proteins / Q000378:metabolism; D015027:Zebrafish; D020666:ras-GRF1 / Q000737:chemistry
D000199:Actins; D048868:Adaptor Proteins, Signal Transducing; D009218:Myosins; D009841:Oligonucleotides; D025521:Tumor Suppressor Proteins; D033921:Vesicular Transport Proteins; D020666:ras-GRF1; C000620875:Golgin45 protein, mouse; C554819:LURAP1 protein, human; C000621004:Lrap35a protein, mouse; C513037:Rasgrf1 protein, mouse; C407152:MYO18A protein, human