cdc2 Kinase Phosphorylation of Desmin at Three Serine/Threonine Residues in the Amino-Terminal Head Domain

doi:10.1006/bbrc.1993.1138

Biochemical and Biophysical Research Communications

Volume 190, Issue 3, 15 February 1993, Pages 927-934

https://doi.org/10.1006/bbrc.1993.1138 Get rights and content

Abstract

Phosphorylation of desmin filament by cdc2 kinase Induced a transition toward the depolymerized state of the filament. Sequence analysis of purified phosphopeptides derived from cdc2 kinase-phosphorylated desmin revealed that Ser-6, Ser-22 and Thr-64 in the N-terminal head domain were the sites phosphorylated.

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Diseases of the Cytoskeleton: The Desminopathies
2017, Cardioskeletal Myopathies in Children and Young Adults
The striated muscle cell cytoskeleton is a multifunctional, dynamic, and complex structure that serves as a scaffold to maintain the structural integrity and architecture of the cell. However, it also functions as a platform for inter- and intracellular signaling, linking the contractile apparatus so that sarcomeres may function in concert, and linking those structures in turn to the sarcolemma, cell–cell junctions, the mitochondria, and nucleus. This review focuses on the intermediate filament and associated protein components of the cytoskeleton and explores how mutations in desmin lead to cardiomyopathy. The clinical and pathological features of these myopathies are described with a particular focus on placing these events within the broader context of proteotoxicity and its relevance as a converging pathway in cardiac disease.
Desmin phosphorylation by Cdk1 is required for efficient separation of desmin intermediate filaments in mitosis and detected in murine embryonic/newborn muscle and human rhabdomyosarcoma tissues
2016, Biochemical and Biophysical Research Communications
Citation Excerpt :
These results confirmed our previous report that desmin phosphorylation by Cdk1 induced the disassembly of desmin-IFs in vitro [30]. We previously reported that Cdk1 can phosphorylate Ser6, Ser22, and Thr64 on chicken desmin [30]. According to a public database PhosphoSitePlus® (http://www.phosphosite.org), Ser6 and Ser22 on chicken desmin are corresponding to Ser6 and Ser31 on murine desmin, respectively; the first Met is uncounted in each amino acid position.
Desmin is a type III intermediate filament (IF) component protein expressed specifically in muscular cells. Desmin is phosphorylated by Aurora-B and Rho-kinase specifically at the cleavage furrow from anaphase to telophase. The disturbance of this phosphorylation results in the formation of unusual long bridge-like IF structures (IF-bridge) between two post-mitotic (daughter) cells. Here, we report that desmin also serves as an excellent substrate for the other type of mitotic kinase, Cdk1. Desmin phosphorylation by Cdk1 loses its ability to form IFs in vitro. We have identified Ser6, Ser27, and Ser31 on murine desmin as phosphorylation sites for Cdk1. Using a site- and phosphorylation-state-specific antibody for Ser31 on desmin, we have demonstrated that Cdk1 phosphorylates desmin in entire cytoplasm from prometaphase to metaphase. Desmin mutations at Cdk1 sites exhibit IF-bridge phenotype, the frequency of which is significantly increased by the addition of Aurora-B and Rho-kinase site mutations to Cdk1 site mutations. In addition, Cdk1-induced desmin phosphorylation is detected in mitotic muscular cells of murine embryonic/newborn muscles and human rhabdomyosarcoma specimens. Therefore, Cdk1-induced desmin phosphorylation is required for efficient separation of desmin-IFs and generally detected in muscular mitotic cells in vivo.
Identification of substrates for cyclin dependent kinases
2010, Advances in Enzyme Regulation
Differentiation of cardiomyocytes requires functional serine residues within the amino-terminal domain of desmin
2007, Differentiation
Desmin contributes to the stability of the myocardium and its amino-terminal domain influences intermediate filament formation and interacts with a variety of proteins and DNAs. Specific serine residues located in this domain are reversibly phosphorylated in a cell cycle and developmental stage-dependent manner as has been demonstrated also for other cytoplasmic type III intermediate filament proteins. Although absence of desmin apparently does not affect cardiomyogenesis, homozygous deletion of the amino-terminal domain of desmin severely inhibited in vitro cardiomyogenesis. To demonstrate the significance of phosphorylation of this domain in cardiomyogenic commitment and differentiation, we inhibited phosphorylation of serine residues 6, 7, and 8 by mutation to alanine, and investigated early cardiomyogenesis in heterozygous embryoid bodies. As control, serine residues 31 and 32, which are not phosphorylated by kinases mutating serine residues 6, 7, and 8, were mutated to alanine in a second set. Desmin^S6,7,8A interfered with cardiomyogenesis and myofibrillogenesis in a dominant negative fashion, whereas desmin^S31,32A produced only a mild phenotype. Desmin^S6,7,8A led to the down-regulation of the transcription factor genes brachyury, goosecoid, nkx2.5, and mef2C and increased apoptosis of presumptive mesoderm and differentiating cardiomyocytes. Surviving cardiomyocytes which were few in number had no myofibrils. Demonstration that some but not any mutant desmin interfered with the very beginning of cardiomyogenesis suggests an important function of temporarily phosphorylated serine residues 6, 7, and 8 in the amino-terminal domain of desmin in cardiomyogenic commitment and differentiation.
Domain-Specific Phosphorylation as a Regulator of Intermediate Filaments
2006, Advances in Molecular and Cell Biology
Citation Excerpt :
The specific sites found to be responsible for the phosphorylation‐mediated depolymerization were localized on the N‐terminal domain of vimentin (Ando et al., 1989). Subsequently, the preference of N‐terminal phosphorylation in disassembly of IFs and in reduced capability to form filaments in vitro has been demonstrated with several IF proteins, for example, vimentin (Evans, 1988b; Chou et al., 1990; Matsuzawa et al., 1997; Goto et al., 1998, 2002, 2003; Eriksson et al., 2004), desmin (Geisler and Weber, 1988; Kusubata et al., 1993; Inada et al., 1998; Kawajiri et al., 2003), GFAP (Inagaki et al., 1990; Kosako et al., 1997; Kawajiri et al., 2003), and low‐molecular weight neurofilament (NF‐L) (Gonda et al., 1990; Nakamura et al., 1990; Mukai et al., 1996). In further support of the specific role of the N‐terminus in IF assembly/disassembly, studies using surface plasmon resonance have indicated that the N‐terminal domain of vimentin binds to the 2B helix in the rod domain and that phosphorylation interferes with this interaction (Gohara et al., 2001).
Intermediate filaments (IFs) are cytoskeletal structures that maintain cell and tissue integrity. In addition, they have been implicated in a number of other functions, including maintenance of cell shape, subcellular organelle positioning, support to cell migration, radial growth of axons, and scaffolding of signaling molecules. Many of the recognized tasks are affected by phosphorylation, which is the key mechanism to regulate both the organization of IFs and the association of IFs with interacting molecules. Another important function of IF phosphorylation is the disassembly of cytoplasmic and karyoskeletal IF networks during mitosis. An intriguing feature of IFs is the tripartite structure of their building blocks, with a highly conserved central rod domain, which is flanked by variable length N‐ and C‐terminal regions. While the rod domain is crucial for the assembly of IFs, a common feature of the distal domains is that they represent the main targets of phosphorylation. Since phosphate targeting seems to have domain‐specific functions, this chapter aims at summarizing the different roles of N‐ and C‐terminal phosphorylation of IFs.
p21-activated kinase PAK phosphorylates desmin at sites different from those for Rho-associated kinase
2000, Biochemical and Biophysical Research Communications
p21-activated kinase (PAK) and Rho-associated kinase (Rho-kinase) have been shown to induce Ca²⁺-independent contraction of smooth muscle. PAK-induced contraction of Triton-skinned smooth muscle correlates with increased phosphorylation of caldesmon and desmin, although the role of desmin phosphorylation has remained obscure. Here we report that desmin serves as an excellent substrate for PAK in vitro. PAK phosphorylated desmin in a GTP · Cdc42/Rac-dependent manner. Phosphorylation of desmin by PAK dramatically inhibited its filament-forming ability. PAK phosphorylated mainly serine residues of the head domain of desmin, and the major phosphorylation sites differed from those for Rho-kinase. These results suggest that different site-specific phosphorylation of desmin via two divergent protein kinases downstream of Rho family GTPases would seem to increase the regulatory potential for organization of desmin filaments.

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Regular Articlecdc2 Kinase Phosphorylation of Desmin at Three Serine/Threonine Residues in the Amino-Terminal Head Domain

Abstract

Regular Article
cdc2 Kinase Phosphorylation of Desmin at Three Serine/Threonine Residues in the Amino-Terminal Head Domain