Chaperone-mediated reversible inhibition of the sarcomeric myosin power stroke.

Molecular chaperones are required for successful folding and assembly of sarcomeric myosin in skeletal and cardiac muscle. Here, we show that the chaperone UNC-45B inhibits the actin translocation function of myosin. Further, we show that Hsp90, another chaperone involved in sarcomere development, allows the myosin to resume actin translocation.

Overexpression of smooth muscle myosin heavy chain leads to activation of the unfolded protein response and autophagic turnover of thick filament-associated proteins in vascular smooth muscle cells.

Duplications spanning nine genes at the genomic locus 16p13.1 predispose individuals to acute aortic dissections. The most likely candidate gene in this region leading to the predisposition for dissection is MYH11, which encodes smooth muscle myosin heavy chain (SM-MHC).

Paradigm shifts in cardiovascular research from Caenorhabditis elegans muscle.

Research on Caenorhabditis elegans has led to the discovery of the consequences of mutation in myosin, its associated proteins, and the extracellular matrix-membrane cytoskeleton complex. Key results include understanding thick filament structure and assembly, the regulation of sarcomeric protein turnover, and the organization of thick and thin filaments into ordered sarcomeres. These results are critical to studies of cardiovascular diseases such as the cardiomyopathies, congenital septal defects, aneurysms of the thoracic aorta, and cardiac remodeling in heart failure.

Tracking UNC-45 chaperone-myosin interaction with a titin mechanical reporter.

Myosins are molecular motors that convert chemical energy into mechanical work. Allosterically coupling ATP-binding, hydrolysis, and binding/dissociation to actin filaments requires precise and coordinated structural changes that are achieved by the structurally complex myosin motor domain. UNC-45, a member of the UNC-45/Cro1/She4p family of proteins, acts as a chaperone for myosin and is essential for proper folding and assembly of myosin into muscle thick filaments in vivo.

Dual function of the UNC-45b chaperone with myosin and GATA4 in cardiac development.

Cardiac development requires interplay between the regulation of gene expression and the assembly of functional sarcomeric proteins. We report that UNC-45b recessive loss-of-function mutations in C3H and C57BL/6 inbred mouse strains cause arrest of cardiac morphogenesis at the formation of right heart structures and failure of contractile function. Wild-type C3H and C57BL/6 embryos at the same stage, E9.

Caenorhabditis elegans muscle: a genetic and molecular model for protein interactions in the heart.

The nematode Caenorhabditis elegans has become established as a major experimental organism with applications to many biomedical research areas. The body wall muscle cells are a useful model for the study of human cardiomyocytes and their homologous structures and proteins. The ability to readily identify mutations affecting these proteins and structures in C elegans and to be able to rigorously characterize their genotypes and phenotypes at the cellular and molecular levels permits mechanistic studies of the responsible interactions relevant to the inherited human cardiomyopathies.

The myosin-binding UCS domain but not the Hsp90-binding TPR domain of the UNC-45 chaperone is essential for function in Caenorhabditis elegans.

The UNC-45 family of molecular chaperones is expressed in metazoan organisms from Caenorhabditis elegans to humans. The UNC-45 protein is essential in C. elegans for early body-wall muscle cell development and A-band assembly.

Differential turnover of myosin chaperone UNC-45A isoforms increases in metastatic human breast cancer.

UNC-45A is a molecular chaperone targeted to non-muscle myosins and is essential for cell division. Here, we show that UNC-45A mRNA and protein expression was elevated in human breast carcinomas and cell lines derived from breast carcinoma metastases. Moreover, small hairpin RNA knockdowns of endogenously overexpressed UNC-45A in the most metastatic cell line led to significant decreases in the rates of cell proliferation and invasion, concomitant with reduction in the interaction of myosin II with actin filaments.

Drosophila UNC-45 accumulates in embryonic blastoderm and in muscles, and is essential for muscle myosin stability.

UNC-45 is a chaperone that facilitates folding of myosin motor domains. We have used Drosophila melanogaster to investigate the role of UNC-45 in muscle development and function. Drosophila UNC-45 (dUNC-45) is expressed at all developmental stages.

The UNC-45 chaperone mediates sarcomere assembly through myosin degradation in Caenorhabditis elegans.

Myosin motors are central to diverse cellular processes in eukaryotes. Homologues of the myosin chaperone UNC-45 have been implicated in the assembly and function of myosin-containing structures in organisms from fungi to humans. In muscle, the assembly of sarcomeric myosin is regulated to produce stable, uniform thick filaments.

UNC-98 links an integrin-associated complex to thick filaments in Caenorhabditis elegans muscle.

Focal adhesions are multiprotein assemblages that link cells to the extracellular matrix. The transmembrane protein, integrin, is a key component of these structures. In vertebrate muscle, focal adhesion-like structures called costameres attach myofibrils at the periphery of muscle cells to the cell membrane.

Regulation of the myosin-directed chaperone UNC-45 by a novel E3/E4-multiubiquitylation complex in C. elegans.

The organization of the motor protein myosin into motile cellular structures requires precise temporal and spatial control. Caenorhabditis elegans UNC-45 facilitates this by functioning both as a chaperone and as a Hsp90 cochaperone for myosin during thick filament assembly. Consequently, mutations in C.

Homodimerization through coiled-coil regions enhances activity of the myotonic dystrophy protein kinase.

Myotonic dystrophy protein kinase (DMPK) is the protein product of the human DM-1 locus on chromosome 19q13.1 and has been implicated in the cardiac and behavioral dysfunctions of the disorder. DMPK contains four distinct regions: a leucine-rich repeat (L), a serine-threonine protein kinase catalytic domain (PK), an alpha-helical coiled-coil region (H), and a putative transmembrane-spanning tail (T).

Novel phosphorylation target in the serum response factor MADS box regulates alpha-actin transcription.

Serum response factor (SRF) is a phosphoprotein that regulates skeletal and cardiac alpha-actin gene transcription. Myotonic dystrophy protein kinase (DMPK), a muscle- and neuron-restricted kinase, enhanced SRF-mediated promoter activity of the skeletal and cardiac alpha-actin genes in C2C12 myoblasts as well as in nonmyogenic cells. DMPK phosphorylated SRF in vitro in the alphaI coil of the DNA-binding domain in the MADS box, a highly conserved region required for DNA binding, dimerization, and co-activator interaction in COS and CV1 cells.

A role for myotonic dystrophy protein kinase in synaptic plasticity.

Myotonic dystrophy (DM) is associated with an expanded triplet repeat in the 3'-untranslated region of the gene for myotonic dystrophy protein kinase (DMPK), which may reduce DMPK expression. It is unclear how reduced DMPK expression might contribute to the symptoms of DM because the normal function of DMPK is not yet understood. Thus we investigated the function of DMPK to gain insight into how reduced DMPK expression might lead to cognitive dysfunction in DM.

Two mammalian UNC-45 isoforms are related to distinct cytoskeletal and muscle-specific functions.

Previous studies have shown that the UNC-45 protein of C. elegans is required for normal thick filament assembly, binds Hsp90 and the myosin head, and shows molecular chaperone activity. We report here that mice and humans each have two genes that are located on different chromosomes, encode distinct UNC-45-like protein isoforms, and are expressed either in multiple tissues or only in cardiac and skeletal muscles.

The UCS family of myosin chaperones.

The canonical UCS (UNC-45/Cro1/She4p) protein, Caenorhabditis elegans UNC-45, was one of the earliest molecules to be shown genetically to be necessary for sarcomere assembly. Genetic analyses of homologues in several fungal species indicate that the conserved UCS domain functionally interacts with conventional type II and unconventional type V myosins. In C.

Role of the myosin assembly protein UNC-45 as a molecular chaperone for myosin.

The organization of myosin into motile cellular structures requires precise temporal and spatial regulation. Proteins containing a UCS (UNC-45/CRO1/She4p) domain are necessary for the incorporation of myosin into the contractile ring during cytokinesis and into thick filaments during muscle development. We report that the carboxyl-terminal regions of UNC-45 bound and exerted chaperone activity on the myosin head.