DOCSLIB.ORG

Posttranslational Regulation of Smads

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Posttranslational Regulation of Smads Downloaded from http://cshperspectives.cshlp.org/ on September 26, 2021 - Published by Cold Spring Harbor Laboratory Press Posttranslational Regulation of Smads Pinglong Xu,1 Xia Lin,2 and Xin-Hua Feng1,2,3 1Life Sciences Institute and Innovation Center for Cell Signaling Network, Zhejiang University, Hangzhou, Zhejiang 310058, China 2Michael E. DeBakey Department of Surgery, Baylor College of Medicine, Houston, Texas 77030 3Department of Molecular & Cellular Biology, Baylor College of Medicine, Houston, Texas 77030 Correspondence: [email protected] Transforming growth factor b (TGF-b) family signaling dictates highly complex programs of gene expression responses, which are extensively regulated at multiple levels and vary depending on the physiological context. The formation, activation, and destruction of two major functional complexes in the TGF-b signaling pathway (i.e., the TGF-b receptor com- plexes and the Smad complexes that act as central mediators of TGF-b signaling) are direct targets for posttranslational regulation. Dysfunction of these complexes often leads or con- tributes to pathogenesis in cancer and fibrosis and in cardiovascular, and autoimmune diseases. Here we discuss recent insights into the roles of posttranslational modifications in the functions of the receptor-activated Smads in the common Smad4 and inhibitory Smads, and in the control of the physiological responses to TGF-b. It is now evident that these modifications act as decisive factors in defining the intensity and versatility of TGF-b responsiveness. Thus, the characterization of posttranslational modifications of Smads not only sheds light on how TGF-b controls physiological and pathological processes but may also guide us to manipulate the TGF-b responses for therapeutic benefits. he transforming growth factor b (TGF-b) main of the receptor upon TGF-b1 binding Tfamily consists of structurally related poly- provides the basis for signal transduction. Li- peptide dimers that are involved in a diversity of gand binding stabilizes the interaction of TGF- physiological processes in all metazoans. They b type II receptor (TbRII) dimers with dimers regulate cell functions, cell and tissue differen- of the TGF-b type I receptor, TbRI, also called tiation, and morphogenesis. As a result, dereg- activin receptor-like kinase 5 (ALK-5), enabling ulation of TGF-b family activities leads to or TbRII to phosphorylate serines and threonines contributes to the pathogenesis of cancer and in the short juxtamembrane Gly-Ser (GS) mo- fibrosis, cardiovascular and autoimmune dis- tifs of TbRI receptors (Souchelnytskyi et al. eases, and developmental defects. 1996). This phosphorylation event triggers con- The signal transduction pathway activated formational changes in TbRI that allow a sur- by TGF-b family proteins appears quite face of TbRI to interact with a basic patch of the straightforward. Viewing TGF-b1 as the proto- Mad homology 2 (MH2) domain of the recep- type, phosphorylation in the cytoplasmic do- tor-regulated Smads (R-Smads) and phosphor- Editors: Rik Derynck and Kohei Miyazono Additional Perspectives on The Biology of the TGF-b Family available at www.cshperspectives.org Copyright # 2016 Cold Spring Harbor Laboratory Press; all rights reserved Advanced Online Article. Cite this article as Cold Spring Harb Perspect Biol doi: 10.1101/cshperspect.a022087 1 Downloaded from http://cshperspectives.cshlp.org/ on September 26, 2021 - Published by Cold Spring Harbor Laboratory Press P. Xu et al. ylation of R-Smads by TbRI on distal two ser- activated in response to stimulation of TGF-b, ines of the conserved carboxy-terminal SSXS nodal, and activin, whereas Smad1, Smad5, motif (Huse et al. 2001; Shi and Massague´ and Smad8 respond to bone-morphogenetic 2003). Subsequently, two phosphorylated R- proteins (BMPs) and growth and differentia- Smads form a heterotrimeric complex with tion factors (GDFs) (Heldin and Moustakas the common Smad4 (co-Smad). The resulting 2012). Characterized by a highly conserved ami- trimeric complex is rapidly translocated into no-terminal MH1 and carboxy-terminal MH2 the nucleus where it elicits transcriptional re- domains, and separated by a variable serine- sponses (Feng and Derynck 2005; Massague´ and proline-rich linker region, these R-Smads et al. 2005; Schmierer and Hill 2007). The act as main effectors for TGF-b family signaling Smad complex associates with a wide range of (Shi and Massague´ 2003). Like many key signal- DNA-binding transcription factors and cofac- ing mediators, the expression level and activity tors (e.g., coactivators and corepressors) to de- of R-Smads are under stringent control, partic- liver diverse transcriptional responses on the ularly through PTMs including (de)phosphor- DNA in a context-dependent manner. Besides ylation, (de)ubiquitylation, sumoylation, (de)- the canonical Smad-dependent pathway, ligand acetylation, methylation, and ADP-ribosylation binding to cognate receptor complexes also (Fig. 1). leads to activation of non-Smad signaling path- ways, such as mitogen-activated protein kinase (MAPK) pathways and the phosphoinositide 3- kinase (PI3K)-Akt-mTOR pathway (Derynck Phosphorylation and Dephosphorylation and Zhang 2003). These non-Smad pathways of R-Smads play an important role in some TGF-b-regulat- Phosphorylation of the SSXS Motif by TGF-b ed processes, such as the generation of regula- Type I Receptor and Other Kinases tory Treg (T) and TH17 cells (Korn et al. 2009). All signaling mediators in the TGF-b path- R-Smads are activated through ligand-induced way are subject to extensive posttranslational phosphorylation by the type I receptors of modifications (PTMs). Taking TGF-b receptors two serines in their carboxy-terminal SSXS mo- as examples, a variety of modifications have tif. The TGF-b-specific type I receptor TbRI/ been identified in recent years. These include ALK-5 phosphorylates Smad2 and Smad3, phosphorylation (Lawler et al. 1997; Galliher whereas the BMP-specific type I receptors and Schiemann 2007; Lee et al. 2007b), ubiqui- ALK-3/BMPRIA (BMP receptor, type IA) and tylation, sumoylation (Kang et al. 2008), ned- ALK-6/BMPRIB phosphorylate Smad1, 5, and dylation (Zuo et al. 2013), and ectodomain 8. This phosphorylation leads to the dissocia- shedding (Atfi et al. 2007; Liu et al. 2009a; Xu tion of R-Smads from their type I receptors, and and Derynck 2010; Mu et al. 2011), which con- homo- or heterodimerization of phosphorylat- tribute to the spatial and temporal regulation in ed R-Smads. After association of two R-Smads TGF-b responsiveness and function in distinct with Smad4, the R-Smad/Smad4 complexes contexts. PTMs of TGF-b receptors have been translocate into the nucleus, in which they elicit reviewed elsewhere (Xu et al. 2012). We now transcriptional responses by interacting with a focus our discussion on the current knowledge variety of transcription factors and transcrip- of the dynamic regulation of distinct Smads by tional coregulators. PTMs, and its pivotal roles in controlling TGF- In addition to this canonical model of TGF- b family functions. b signaling through Smad2 and Smad3, some TGF-b family ligands were shown to initiate unexpected phosphorylation of a noncanonical POSTTRANSLATIONAL MODIFICATIONS R-Smad. Specifically, in addition to the ex- OF R-Smads pected Smad2 and Smad3 activation, TGF-b The eight Smads identified in vertebrates com- induces Smad1 and Smad5 phosphorylation. prise five R-Smads. Smad2 and Smad3 are In endothelial cells, the TGF-b-specific type I 2 Advanced Online Article. Cite this article as Cold Spring Harb Perspect Biol doi: 10.1101/cshperspect.a022087 Downloaded from http://cshperspectives.cshlp.org/ Advanced Online Article. Cite this article as P Phosphorylation SCP1/2/3 Rock U Monoubiquitylation p38 MAPK GSK3 Ac Acetylation MPK38 PP2A ERK p300/CBP MTMR4 USP15 ERK JNK PPM1A Ac SCP1/2/3 Smurf2 CDK8/9 CDK8/9 MPS1 onSeptember26,2021-PublishedbyColdSpringHarborLaboratoryPress CDK2/4 GSK3β NEDD4 CDK2/4 CDK2/4 PKG Smurf2 PKG CK1γ 2 TβR1 Cold Spring Harb Perspect Biol P PPPPPPUUUP T8 T66 K81 T179 S204 S208 S213 S309 K333 K378 S388 S418 S423 S425 Smad3 MH1 Linker MH2 doi: 10.1101 Smad2 MH1 Linker MH2 Posttranslational Regulation of Smads T8 K19 K20 K39 / T197 T220 T255 S110 S240 S245 S250 S260 S417 S465 S467 cshperspect.a022087 PP P Ac PPPP PAK2 TβR1 ERK p300 CaMK II GRK2 ERK CaMK II PAK2 P/CAF CDK8/9 Araf PAK4 ERK PPM1A MPK38 MTMR4 Figure 1. Defined modifications and modifying enzymes of Smad2 and Smad3. Domain structures are indicated. Blue triangles represent modifications that promote Smad signaling, whereas red triangles (upside down) represent modifica- tions that attenuate Smad signaling. 3 Downloaded from http://cshperspectives.cshlp.org/ on September 26, 2021 - Published by Cold Spring Harbor Laboratory Press P. Xu et al. receptor TbRI/ALK-5 as well as the BMP family type I TGF-b family receptor kinases. For exam- type I receptor ALK-1 are essential for TGF- ple, Mps1, a kinase that regulates mitotic pro- b-induced Smad1 and Smad5 phosphoryla- gression, can phosphorylate the SSXS motif tion, together with the accessory receptor en- of Smad2 and Smad3, although it is unclear doglin (Goumans et al. 2002, 2003; Lebrin whether it activates the Smad pathway (Zhu et al. 2004). Although TbRI/ALK-5-mediated et al. 2007). The oncogenic kinase p21 pro- Smad2 and Smad3 activation leads to inhibition tein-activated kinase 4 (PAK4) can phosphory- of cell migration and proliferation, ALK-1-me- late Ser465 in the SSXS motif of Smad2, which diated Smad1 and Smad5 activation results in leads to Smad2 proteasomal degradation. Ac- opposite effects. Therefore, a fine balance be- cordingly, a marked increase in phospho-S465 tween TbRI/ALK-5 and ALK-1 signaling de- Smad2 level, concomitant with a lower total fines the roles of TGF-b in endothelium (Gou- Smad2 protein level, has been observed in gas- mans et al. 2002, 2003; Lebrin et al. 2004). A tric cancer tissues (Wang et al. 2014a). The similar scenario is also observed in dermal fi- physiological significance of R-Smad phos- broblasts, in which TGF-b stimulates Smad1 phorylation and activation by these kinases re- phosphorylation and Smad1-mediated tran- mains to be further explored.
Load more

Recommended publications
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • Pathways of Cellular Proteostasis in Aging and Disease
    JCB: Review Pathways of cellular proteostasis in aging and disease Courtney L. Klaips, Gopal Gunanathan Jayaraj, and F. Ulrich Hartl Department of Cellular Biochemistry, Max Planck Institute of Biochemistry, Martinsried, Germany Ensuring cellular protein homeostasis, or proteostasis, re- control of abundance and subcellular localization, and finally, quires precise control of protein synthesis, folding, confor- disposal by degradation. A complex proteostasis network (PN) acts at each of these mational maintenance, and degradation. A complex and steps to maintain a balanced proteome linked by molecular adaptive proteostasis network coordinates these processes chaperones of different classes as central players. These factors with molecular chaperones of different classes and their ensure de novo folding in a crowded cellular environment and regulators functioning as major players. This network maintain proteins in a soluble, nonaggregated state. Moreover, in conditions that disfavor folding or solubility, certain chaper- serves to ensure that cells have the proteins they need ones act to target misfolded proteins for degradation or spatial while minimizing misfolding or aggregation events that sequestration, thus protecting the rest of the proteome from ab- are hallmarks of age-associated proteinopathies, includ- errant interactions (Balchin et al., 2016; Sontag et al., 2017). Here, we describe the major pathways of cellular pro- ing neurodegenerative disorders such as Alzheimer’s and teostasis and outline the challenges they face during aging and Parkinson’s diseases. It is now clear that the capacity of disease. We exemplify these processes using mainly the proteo- cells to maintain proteostasis undergoes a decline during stasis pathways operating in the cytosol, where most cellular aging, rendering the organism susceptible to these pa- proteins are produced.
    [Show full text]
  • Deubiquitylases in Developmental Ubiquitin Signaling and Congenital Diseases
    Cell Death & Differentiation (2021) 28:538–556 https://doi.org/10.1038/s41418-020-00697-5 REVIEW ARTICLE Deubiquitylases in developmental ubiquitin signaling and congenital diseases 1 1,2 1 Mohammed A. Basar ● David B. Beck ● Achim Werner Received: 16 October 2020 / Revised: 20 November 2020 / Accepted: 24 November 2020 / Published online: 17 December 2020 This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply 2020 Abstract Metazoan development from a one-cell zygote to a fully formed organism requires complex cellular differentiation and communication pathways. To coordinate these processes, embryos frequently encode signaling information with the small protein modifier ubiquitin, which is typically attached to lysine residues within substrates. During ubiquitin signaling, a three-step enzymatic cascade modifies specific substrates with topologically unique ubiquitin modifications, which mediate changes in the substrate’s stability, activity, localization, or interacting proteins. Ubiquitin signaling is critically regulated by deubiquitylases (DUBs), a class of ~100 human enzymes that oppose the conjugation of ubiquitin. DUBs control many essential cellular functions and various aspects of human physiology and development. Recent genetic studies have fi 1234567890();,: 1234567890();,: identi ed mutations in several DUBs that cause developmental disorders. Here we review principles controlling DUB activity and substrate recruitment that allow these enzymes to regulate ubiquitin signaling during development. We summarize key mechanisms of how DUBs control embryonic and postnatal differentiation processes, highlight developmental disorders that are caused by mutations in particular DUB members, and describe our current understanding of how these mutations disrupt development. Finally, we discuss how emerging tools from human disease genetics will enable the identification and study of novel congenital disease-causing DUBs.
    [Show full text]
  • Protein Homeostasis, Second Edition
    This is a free sample of content from Protein Homeostasis, Second Edition. Click here for more information on how to buy the book. Index A overview of diseases, 13, 41–43 Abf2, 160 polyphosphate AD. See Alzheimer’s disease amyloidogenic protein interactions, 393–395 AFG3L2, 162 cytotoxicity amelioration, 396–397 Africa swine fever virus (ASFV), 468 prospects for study, 52, 55 Aging protein homeostasis context, 48–49 heat shock factors, 431 structure–activity relationship, 14–15 proteostasis network regulation, 450–452 therapeutic intervention, 50–52 Ago2, 335, 467 toxicity, 47–48 Aha1, 333, 335, 338–339, 504 Amyotrophic lateral sclerosis (ALS), 214, 262, 264, AIRAP, 267 290–291, 293, 404, 409, 524, 532 AIRAPL, 267–268 ApoB, 135 AKT, 274, 467 ASFV. See Africa swine fever virus ALK, 503 ATAD1, 104 ALMC2, 487 ATF4, 79, 88, 182, 186, 189, 445–446 ALP. See Autophagy lysosome pathway ATF5, 182, 186, 189 a-Synuclein, 375, 522 ATF6, 60, 62–64, 79, 445–446, 481–482, 484–489 ALS. See Amyotrophic lateral sclerosis ATFS-1, 99, 180–181, 185, 188–189, 445–446 Alzheimer’s disease (AD), 186, 519 Atg7, 293 autophagy lysosomal pathway, 290–291 Atg32, 167 chaperone studies, 213–214, 216–217, 222 ATM, 313 Hsp90 studies, 337 ATR, 313 polyphosphate studies, 392, 394–395, 399 Atropine, 501 AMPK, 444 ATXN2, 524 Amyloid Autophagy lysosome pathway (ALP) chaperone modulation chaperone-mediated autophagy, 288–289 aggregation prevention, 217–222 history of study, 285 degradation of amyloid-forming proteins, 225–227 macroautophagy, 287–288 disaggregation of amyloids,
    [Show full text]
  • Functional Modules of the Proteostasis Network
    Downloaded from http://cshperspectives.cshlp.org/ on September 28, 2021 - Published by Cold Spring Harbor Laboratory Press Functional Modules of the Proteostasis Network Gopal G. Jayaraj, Mark S. Hipp, and F. Ulrich Hartl Department of Cellular Biochemistry, Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany Correspondence: [email protected] Cells invest in an extensive network of factors to maintain protein homeostasis (proteostasis) and prevent the accumulation of potentially toxic protein aggregates. This proteostasis network (PN) comprises the machineries for the biogenesis, folding, conformational mainte- nance, and degradation of proteins with molecular chaperones as central coordinators. Here, we review recent progress in understanding the modular architecture of the PN in mammalian cells and how it is modified during cell differentiation. We discuss the capacity and limitations of the PN in maintaining proteome integrity in the face of proteotoxic stresses, such as aggregate formation in neurodegenerative diseases. Finally, we outline various pharmaco- logical interventions to ameliorate proteostasis imbalance. roteins are the most versatile macromole- 1999; Hartl and Hayer-Hartl 2009). However, Pcules and responsible for almost all cellular proteins with complex domain folds and multi- functions. An average human cell expresses domain proteins, which make up the major part ∼10,000–13,000 different protein species (Bek- of the proteome, frequently populate folding in- ker-Jensen et al. 2017; Kulak et al. 2017) with termediates that expose hydrophobic amino acid copy numbers varying over several orders of residues. These proteins are at risk of misfolding magnitude, from afew molecules to tens of thou- and aggregation within the highly crowded en- sands.
    [Show full text]
  • UBE2O Is a Quality Control Factor for Orphans of Multi-Protein Complexes
    RESEARCH ◥ factors. A transmembrane (TM) domain inter- REPORT rupted by three basic residues (hereafter termed 3R) (fig. S1A) is not recognized by quality control factors specialized for mislocalized membrane PROTEOSTASIS proteins, such as BAG6 or the ubiquilin family members (5–7); yet, 3R was ubiquitinated sim- ilarly to a BAG6 substrate (fig. S1B). Although 3R UBE2O is a quality control factor for is an artificial mutant, we reasoned that mech- anistic dissection of its ubiquitination pathway orphans of multiprotein complexes mightleadustoaqualitycontrolpathwayand provide tools that could be exploited to identify physiological substrates. † Kota Yanagitani,* Szymon Juszkiewicz, Ramanujan S. Hegde In vitro–translated 35S-labeled 3R immunopu- rified under native conditions became modified Many nascent proteins are assembled into multiprotein complexes of defined stoichiometry. with His-ubiquitin in the presence of recombinant Imbalances in the synthesis of individual subunits result in orphans. How orphans are selectively E1, E2 (UBCH5), and adenosine 5′-triphosphate eliminated to maintain protein homeostasis is poorly understood. Here, we found that the (ATP) (fig. S1, C and D), indicating that the im- conserved ubiquitin-conjugating enzyme UBE2O directly recognized juxtaposed basic and munopurified complexes contained a ubiquitin hydrophobic patches on unassembled proteins to mediate ubiquitination without a separate ligase. The primary 3R-associated ubiquitination a ubiquitin ligase. In reticulocytes, where UBE2O is highly up-regulated, unassembled -globin activity had a native molecular weight of ~150 to b molecules that failed to assemble with -globin were selectively ubiquitinated by UBE2O. In 300 kD (Fig. 1, A and B). Cross-linking reactions Downloaded from nonreticulocytes, ribosomal proteins that did not engage nuclear import factors were targets for of the active fractions revealed interacting part- UBE2O.
    [Show full text]
  • VIEW Open Access the Role of Ubiquitination and Deubiquitination in Cancer Metabolism Tianshui Sun1, Zhuonan Liu2 and Qing Yang1*
    Sun et al. Molecular Cancer (2020) 19:146 https://doi.org/10.1186/s12943-020-01262-x REVIEW Open Access The role of ubiquitination and deubiquitination in cancer metabolism Tianshui Sun1, Zhuonan Liu2 and Qing Yang1* Abstract Metabolic reprogramming, including enhanced biosynthesis of macromolecules, altered energy metabolism, and maintenance of redox homeostasis, is considered a hallmark of cancer, sustaining cancer cell growth. Multiple signaling pathways, transcription factors and metabolic enzymes participate in the modulation of cancer metabolism and thus, metabolic reprogramming is a highly complex process. Recent studies have observed that ubiquitination and deubiquitination are involved in the regulation of metabolic reprogramming in cancer cells. As one of the most important type of post-translational modifications, ubiquitination is a multistep enzymatic process, involved in diverse cellular biological activities. Dysregulation of ubiquitination and deubiquitination contributes to various disease, including cancer. Here, we discuss the role of ubiquitination and deubiquitination in the regulation of cancer metabolism, which is aimed at highlighting the importance of this post-translational modification in metabolic reprogramming and supporting the development of new therapeutic approaches for cancer treatment. Keywords: Ubiquitination, Deubiquitination, Cancer, Metabolic reprogramming Background cells have aroused increasing attention and interest [3]. Metabolic pathways are of vital importance in proliferat- Because of the generality of metabolic alterations in can- ing cells to meet their demands of various macromole- cer cells, metabolic reprogramming is thought as hall- cules and energy [1]. Compared with normal cells, mark of cancer, providing basis for tumor diagnosis and cancer cells own malignant properties, such as increased treatment [1]. For instance, the application of 18F- proliferation rate, and reside in environments short of deoxyglucose positron emission tomography is based on oxygen and nutrient.
    [Show full text]
  • Post-Translational Modifications of the Energy Guardian AMP-Activated
    International Journal of Molecular Sciences Review Post-Translational Modifications of the Energy Guardian AMP-Activated Protein Kinase Ashley J. Ovens 1,2 , John W. Scott 2,3,4 , Christopher G. Langendorf 3, Bruce E. Kemp 2,3, Jonathan S. Oakhill 1,2 and William J. Smiles 1,* 1 Metabolic Signalling Laboratory, St Vincent’s Institute of Medical Research, School of Medicine, University of Melbourne, Fitzroy, VIC 3065, Australia; [email protected] (A.J.O.); [email protected] (J.S.O.) 2 Mary MacKillop Institute for Health Research, Australian Catholic University, Fitzroy, VIC 3000, Australia; [email protected] (J.W.S.); [email protected] (B.E.K.) 3 Protein Chemistry & Metabolism, St Vincent’s Institute of Medical Research, School of Medicine, University of Melbourne, Fitzroy, VIC 3065, Australia; [email protected] 4 The Florey Institute of Neuroscience and Mental Health, Parkville, VIC 3052, Australia * Correspondence: [email protected] Abstract: Physical exercise elicits physiological metabolic perturbations such as energetic and ox- idative stress; however, a diverse range of cellular processes are stimulated in response to combat these challenges and maintain cellular energy homeostasis. AMP-activated protein kinase (AMPK) is a highly conserved enzyme that acts as a metabolic fuel sensor and is central to this adaptive response to exercise. The complexity of AMPK’s role in modulating a range of cellular signalling cascades is well documented, yet aside from its well-characterised regulation by activation loop phosphorylation, AMPK is further subject to a multitude of additional regulatory stimuli. There- fore, in this review we comprehensively outline current knowledge around the post-translational Citation: Ovens, A.J; Scott, J.W; modifications of AMPK, including novel phosphorylation sites, as well as underappreciated roles for Langendorf, C.G; Kemp, B.E; Oakhill, ubiquitination, sumoylation, acetylation, methylation and oxidation.
    [Show full text]
  • UBE2O Negatively Regulates TRAF6-Mediated NF-Κb Activation by Inhibiting TRAF6 Polyubiquitination
    npg UBE2O inhibits TRAF6-mediated NF-κB signaling Cell Research (2013) 23:366-377. 366 © 2013 IBCB, SIBS, CAS All rights reserved 1001-0602/13 $ 32.00 npg ORIGINAL ARTICLE www.nature.com/cr UBE2O negatively regulates TRAF6-mediated NF-κB activation by inhibiting TRAF6 polyubiquitination Xiaofei Zhang1, *, Juan Zhang1, *, Long Zhang1, Hans van Dam1, Peter ten Dijke1 1Department of Molecular Cell Biology, Cancer Genomics Centre Netherlands and Centre for Biomedical Genetics, Leiden Uni- versity Medical Center, Postbus 9600, 2300 RC Leiden, The Netherlands Tumor necrosis factor (TNF) receptor-associated factor 6 (TRAF6) is a key regulator of the activation of transcrip- tion factor NF-κB by the interleukin-1 receptor (IL-1R)/Toll-like receptor (TLR) superfamily. Recruitment of TRAF6 to the receptor-associated IRAK1-IRAK4-MyD88 adaptor protein complex induces lysine 63 (K63) autopolyubiq- uitination of TRAF6, which leads to further recruitment of downstream regulators, such as TAB2/3 and TAK1, and subsequently triggers NF-κB activation. Here, we identified the putative E2 ubiquitin-conjugating (UBC) enzyme UBE2O as a novel negative regulator of TRAF6-dependent NF-κB signaling. We found that UBE2O binds to TRAF6 to inhibit its K63-polyubiquitination, and to prevent the activation of NF-κB by IL-1β and lipopolysaccharides (LPS). We further show that the inhibitory effect of UBE2O is independent of its carboxy-terminal UBC domain. In con- trast, we found that UBE2O acts to disrupt the IL-1β-induced association of TRAF6 with MyD88. These results pro- vide novel insight into the regulation of signaling by IL-1R/TLR and TRAF6.
    [Show full text]
  • It's Not Just a Phase; Ubiquitination in Cytosolic Protein Quality Control
    Biochemical Society Transactions (2021) 49 365–377 https://doi.org/10.1042/BST20200694 Review Article It’s not just a phase; ubiquitination in cytosolic protein quality control Heather A. Baker1 and Jonathan P. Bernardini1,2,3 1Michael Smith Laboratories, University of British Columbia, Vancouver, Canada; 2Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia; 3Department of Medical Biology, The University of Melbourne, Melbourne, Australia Correspondence: Jonathan P. Bernardini ([email protected]) Downloaded from http://portlandpress.com/biochemsoctrans/article-pdf/49/1/365/905078/bst-2020-0694c.pdf by guest on 27 September 2021 The accumulation of misfolded proteins is associated with numerous degenerative condi- tions, cancers and genetic diseases. These pathological imbalances in protein homeosta- sis (termed proteostasis), result from the improper triage and disposal of damaged and defective proteins from the cell. The ubiquitin-proteasome system is a key pathway for the molecular control of misfolded cytosolic proteins, co-opting a cascade of ubiquitin ligases to direct terminally damaged proteins to the proteasome via modification with chains of the small protein, ubiquitin. Despite the evidence for ubiquitination in this crit- ical pathway, the precise complement of ubiquitin ligases and deubiquitinases that modu- late this process remains under investigation. Whilst chaperones act as the first line of defence against protein misfolding, the ubiquitination machinery has a pivotal role in tar- geting terminally defunct cytosolic proteins for destruction. Recent work points to a complex assemblage of chaperones, ubiquitination machinery and subcellular quarantine as components of the cellular arsenal against proteinopathies. In this review, we examine the contribution of these pathways and cellular compartments to the maintenance of the cytosolic proteome.
    [Show full text]
  • Functional Modules of the Proteostasis Network
    Functional Modules of the Proteostasis Network Gopal G. Jayaraj, Mark S. Hipp and F. Ulrich Hartl Department of Cellular Biochemistry, Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany 1 Abstract Cells invest in an extensive network of factors to maintain protein homeostasis (proteostasis) and prevent the accumulation of potentially toxic protein aggregates. This proteostasis network (PN) comprises the machineries for the biogenesis, folding, conformational maintenance and degradation of proteins, with molecular chaperones as central coordinators. Here we review recent progress in understanding the modular architecture of the PN in mammalian cells and how it is modified during cell differentiation. We discuss the capacity and limitations of the PN in maintaining proteome integrity in the face of proteotoxic stresses, such as aggregate formation in neurodegenerative diseases. Finally, we outline various pharmacological interventions to ameliorate proteostasis imbalance. 2 Introduction Proteins are the most versatile macromolecules and are responsible for almost all cellular functions. An average human cell expresses approximately 10,000- 13,000 different protein species (Bekker-Jensen et al. 2017; Kulak et al. 2017), with copy numbers varying over several orders of magnitude, from a few molecules to tens of thousands. To maintain protein homeostasis, or proteostasis (Balchin et al. 2016), cells must ensure that these proteins fold and assemble correctly and exist in the right cellular locale at appropriate abundance. Most proteins must adopt a unique, thermodynamically stable three-dimensional structure, the functionally active state, that is determined by the amino acid sequence (Anfinsen 1973). Proteins must fold to this native state at biologically relevant timescales, a process that is increasingly well understood, at least for small model proteins that fold rapidly without detectable intermediate states (Dobson and Karplus 1999; Hartl and Hayer-Hartl 2009).
    [Show full text]
  • The HIV-1 Tat Protein Recruits a Ubiquitin Ligase to Reorganize The
    RESEARCH ARTICLE The HIV-1 Tat protein recruits a ubiquitin ligase to reorganize the 7SK snRNP for transcriptional activation Tyler B Faust1, Yang Li1, Curtis W Bacon2, Gwendolyn M Jang3,4, Amit Weiss1, Bhargavi Jayaraman1, Billy W Newton3,4, Nevan J Krogan3,4, Iva´ n D’Orso2, Alan D Frankel1* 1Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United States; 2Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, United States; 3Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States; 4J David Gladstone Institutes, San Francisco, United States Abstract The HIV-1 Tat protein hijacks P-TEFb kinase to activate paused RNA polymerase II (RNAP II) at the viral promoter. Tat binds additional host factors, but it is unclear how they regulate RNAP II elongation. Here, we identify the cytoplasmic ubiquitin ligase UBE2O as critical for Tat transcriptional activity. Tat hijacks UBE2O to ubiquitinate the P-TEFb kinase inhibitor HEXIM1 of the 7SK snRNP, a fraction of which also resides in the cytoplasm bound to P-TEFb. HEXIM1 ubiquitination sequesters it in the cytoplasm and releases P-TEFb from the inhibitory 7SK complex. Free P-TEFb then becomes enriched in chromatin, a process that is also stimulated by treating cells with a CDK9 inhibitor. Finally, we demonstrate that UBE2O is critical for P-TEFb recruitment to the HIV-1 promoter. Together, the data support a unique model of elongation control where non- degradative ubiquitination of nuclear and cytoplasmic 7SK snRNP pools increases P-TEFb levels for transcriptional activation. DOI: https://doi.org/10.7554/eLife.31879.001 *For correspondence: [email protected] Competing interests: The Introduction authors declare that no The advent of genome-wide methods to interrogate transcription has shown that many genes in competing interests exist.
    [Show full text]