Smyd2 controls cytoplasmic lysine methylation of Hsp90 and myofilament organization

Donlin, Laura T.; Andresen, Christian; Just, Steffen; Rudensky, Eugene; Pappas, Christopher T.; Krüger, Martina; Jacobs, Erica Y.; Unger, Andreas; Zieseniß, Anke; Dobenecker, Marc‐Werner; Voelkel, Tobias; Chait, Brian T.; Gregorio, Carol C.; Rottbauer, Wolfgang; Tarakhovsky, Alexander; Linke, Wolfgang A.

doi:10.1101/gad.177758.111

Cited by 141 publications

(193 citation statements)

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“…These include K161me1 on Heterogeneous nuclear ribonucleoprotein D-like, K53me1 on 60S ribosomal protein L36a-like, K1162me3 on Wiz and K737me1 on Heat shock protein 90-α (HSP90α). 12,25,26 HSP90α K737 is known to be monomethylated by methyltransferase SMYD2. 25 The methylation at this site contributed to the formation of SMYD2 protein complex, so as to alter muscle function in muscle cells.…”

Section: Resultsmentioning

confidence: 99%

“…12,25,26 HSP90α K737 is known to be monomethylated by methyltransferase SMYD2. 25 The methylation at this site contributed to the formation of SMYD2 protein complex, so as to alter muscle function in muscle cells. Apart from this known site, a novel dimethylation site at HSP90α K615 was detected, suggesting there might be another biological mechanism to regulate HSP90α through lysine methylation.…”

Section: Resultsmentioning

confidence: 99%

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Large-scale global identification of protein lysine methylation in vivo

Arnaudo²,

2013

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Large-scale global identification of protein lysine methylation in vivo

Arnaudo²,

2013

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“…Importantly, only the methylated HSP90 can interact with N2-A (Figure 3). 55 The Smyd2-methyl-HSP90 complex stabilizes N2-A and helps maintain the sarcomeric I-band structure. Knockdown of Smyd2 in zebrafish specifically disrupts the Z-/I-bands and impairs muscle and heart function.…”

Section: Titin Functions Acquired Through Ligand Bindingmentioning

confidence: 99%

“…Knockdown of Smyd2 in zebrafish specifically disrupts the Z-/I-bands and impairs muscle and heart function. 55,56 To elucidate the conformational changes in methyl-HSP90, which presumably trigger the binding to N2-A, an atomic structure of the complex with Smyd2 would be useful.…”

Section: Titin Functions Acquired Through Ligand Bindingmentioning

confidence: 99%

Gigantic Business

Linke

Hamdani

2014

Circulation Research

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With every heartbeat, our cardiomyocytes are exposed to variable degrees of stress and strain of both internal and external origin. The unique mechanical properties of these myocytes are determined by the sarcomeric cytoskeleton. The actin (thin) and myosin (thick) filaments of the sarcomere are mainly relevant for active force generation, whereas the titin filaments determine sarcomeric viscoelasticity. The giant protein titin, which is the focus of this review, has various roles beyond viscoelastic force generation 1-3 : (1) it keeps the thick filaments centered in the sarcomere, allowing optimum active force development; (2) it is important for the assembly of the sarcomeres; (3) it participates in mechano-chemical signaling events via some of its binding partners; and (4) it may be crucial for the length-dependent activation of the contractile apparatus, which underlies the Frank-Starling law. During the past decade, we have learned that titin elasticity is highly variable in the developing and the adult healthy heart and that it can be pathologically altered in heart disease. These alterations greatly affect the extensibility and the diastolic passive stiffness of myocardium and presumably also the mechanosensitivity. Moreover, TTN, which encodes the titin protein, has been recognized as a major human disease gene. In this context, the properties and functions of cardiac titin have become of interest in the continuing search for the molecular origins of human heart disease and the identification of novel potential targets for therapeutic intervention. In our review, we begin with a short clinical perspective establishing the link between diastolic stiffness and titin and briefly compare the importance of titin versus collagen for myocardial passive stiffness. We then cover some details of titin protein structure and elasticity, before summarizing how titin-binding partners affect titin properties and broaden the range of titin functions, with a special focus on the standing of titin in pathways of protein quality control. We continue with a current overview of titin mutations in human skeletal muscle and heart disease. The remainder of the review deals with the contribution of titin to diastolic stiffness and how it is modulated in normal and failing hearts by mechanisms such as titin-isoform switching, titin phosphorylation (including heart failure [HF]-related alterations of it), and oxidative modifications. Clinical Context: Diastolic Function and Diastolic AbnormalitiesDiastolic function has moved into the focus of HF research, because an increasing number of patients with HF (≥50%) present with diastolic rather than systolic dysfunction. 4 Common abnormalities of diastolic function include impaired left ventricular (LV) relaxation, decreased LV distensibility, increased LV end-diastolic stiffness, and pericardial and right ventricular constraint. These abnormalities have been suggested to be contributing factors in diastolic HF or HF with a preserved ejection fraction (HFpEF).5 HFpEF is accompanied by ...

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“…In addition to substrates identified using biochemical approaches, an investigation into proteins that stably physically associate with SMYD2 led to the discovery of HSP90AA1-K615me1 as a Kme1 site in cells directly regulated by SMYD2 activity (31,32). Despite the identification of potential substrates, the cellular and molecular biology of SMYD2 in oncogenesis remains largely understood-due, at least in part, to a lack of comprehensive characterization of endogenous SMYD2 activity in cells.…”

mentioning

confidence: 99%

Quantitative Profiling of the Activity of Protein Lysine Methyltransferase SMYD2 Using SILAC-Based Proteomics

Olsen

Cao

Han

et al. 2016

Molecular & Cellular Proteomics

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The significance of non-histone lysine methylation in cell biology and human disease is an emerging area of research exploration. The development of small molecule inhibitors that selectively and potently target enzymes that catalyze the addition of methyl-groups to lysine residues, such as the protein lysine mono-methyltransferase SMYD2, is an active area of drug discovery. Critical to the accurate assessment of biological function is the ability to identify target enzyme substrates and to define enzyme substrate specificity within the context of the cell. Here, using stable isotopic labeling with amino acids in cell culture (SILAC) coupled with immunoaffinity enrichment of mono-methyl-lysine (Kme1) peptides and mass spectrometry, we report a comprehensive, large-scale proteomic study of lysine mono-methylation, comprising a total of 1032 Kme1 sites in esophageal squamous cell carcinoma (ESCC) cells and 1861 Kme1 sites in ESCC cells overexpressing SMYD2. Among these Kme1 sites is a subset of 35 found to be potently down-regulated by both shRNA-mediated knockdown of SMYD2 and LLY-507, a selective small molecule inhibitor of SMYD2. In addition, we report specific protein sequence motifs enriched in Kme1 sites that are directly regulated by endogenous SMYD2 activity, revealing that SMYD2 substrate specificity is more diverse than expected. We further show direct activity of SMYD2 toward BTF3-K2, PDAP1-K126 as well as numerous sites within the repetitive units of two unique and exceptionally large proteins, AHNAK and AHNAK2. Collectively, our findings provide quantitative insights into the cellular activity and substrate recognition of SMYD2 as well as the global landscape and regulation of protein mono-methylation. Protein lysine methyltransferases (PKMTs) 1 catalyze the sequence-specific transfer of one, two, or three methyl groups to the side chains of lysine residues (1-4). In addition to the extensively studied lysine methylation on histones, PKMTs can modify non-histone proteins (3,(5)(6)(7)(8)). An increasing number of non-histone proteins have been reported as PKMT substrates, and, as a result, potential roles for the dysregulation of non-histone lysine methylation in cancer development and progression have been proposed (9). The majority of PKMT substrates have been identified based on biochemical methylation assays using recombinant enzyme and substrate, followed by cell-based assays typically requiring overexpression of enzyme and/or substrate to maximize signal detection (10 -13). However, it is difficult to discern whether these substrates are bona fide physiological substrates of endogenous PKMT activity. With the development of PKMT-targeted drugs emerging as a key area of drug discovery (14 -18), unbiased and quantitative methods enabling the comprehensive identification of histone and non-histone substrates in cells are critical to clarifying the cell-relevant substrates of these enzymes and to more accurately understand the functions of non-histone methylation.Progressing from targeted biochemical...

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Smyd2 controls cytoplasmic lysine methylation of Hsp90 and myofilament organization

Cited by 141 publications

References 38 publications

Large-scale global identification of protein lysine methylation in vivo

Large-scale global identification of protein lysine methylation in vivo

Gigantic Business

Quantitative Profiling of the Activity of Protein Lysine Methyltransferase SMYD2 Using SILAC-Based Proteomics

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