Functional interactions between ubiquitin E2 enzymes and TRIM proteins

2011 ◽  
Vol 434 (2) ◽  
pp. 309-319 ◽  
Author(s):  
Luisa M. Napolitano ◽  
Ellis G. Jaffray ◽  
Ronald T. Hay ◽  
Germana Meroni
Keyword(s):  
Coiled Coil ◽  
Specific Interactions ◽  
E3 Ligases ◽  
Cellular Processes ◽  
Coiled Coil Region ◽  
Physiological Relevance ◽  
Tripartite Motif ◽  
E2 Enzymes ◽  
Specific Reactions ◽  
Trim Proteins

The TRIM (tripartite motif) family of proteins is characterized by the presence of the tripartite motif module, composed of a RING domain, one or two B-box domains and a coiled-coil region. TRIM proteins are involved in many cellular processes and represent the largest subfamily of RING-containing putative ubiquitin E3 ligases. Whereas their role as E3 ubiquitin ligases has been presumed, and in several cases established, little is known about their specific interactions with the ubiquitin-conjugating E2 enzymes or UBE2s. In the present paper, we report a thorough screening of interactions between the TRIM and UBE2 families. We found a general preference of the TRIM proteins for the D and E classes of UBE2 enzymes, but we also revealed very specific interactions between TRIM9 and UBE2G2, and TRIM32 and UBE2V1/2. Furthermore, we demonstrated that the TRIM E3 activity is only manifest with the UBE2 with which they interact. For most specific interactions, we could also observe subcellular co-localization of the TRIM involved and its cognate UBE2 enzyme, suggesting that the specific selection of TRIM–UBE2 pairs has physiological relevance. Our findings represent the basis for future studies on the specific reactions catalysed by the TRIM E3 ligases to determine the fate of their targets.

scholarly journals TRIM Proteins and Their Roles in Antiviral Host Defenses

2018 ◽  
Vol 5 (1) ◽  
pp. 385-405 ◽  
Author(s):  
Michiel van Gent ◽  
Konstantin M.J. Sparrer ◽  
Michaela U. Gack
Keyword(s):  
Host Defense ◽  
Host Immunity ◽  
Innate Immune ◽  
Host Defenses ◽  
E3 Ligases ◽  
Cellular Processes ◽  
Cytokine Responses ◽  
Tripartite Motif ◽  
Trim Proteins

Tripartite motif (TRIM) proteins are a versatile family of ubiquitin E3 ligases involved in a multitude of cellular processes. Studies in recent years have demonstrated that many TRIM proteins play central roles in the host defense against viral infection. While some TRIM proteins directly antagonize distinct steps in the viral life cycle, others regulate signal transduction pathways induced by innate immune sensors, thereby modulating antiviral cytokine responses. Furthermore, TRIM proteins have been implicated in virus-induced autophagy and autophagy-mediated viral clearance. Given the important role of TRIM proteins in antiviral restriction, it is not surprising that several viruses have evolved effective maneuvers to neutralize the antiviral action of specific TRIM proteins. Here, we describe the major antiviral mechanisms of TRIM proteins as well as viral strategies to escape TRIM-mediated host immunity.

Abstract 218: The Role of Novel Tripartite Motif Proteins in Sarcolemmal Membrane Repair

2014 ◽  
Vol 115 (suppl_1) ◽  
Author(s):  
Liubov V Gushchina ◽  
Jenna Alloush ◽  
Sayak Bhattacharya ◽  
Zhaobin Xu ◽  
Eric X Beck ◽  
...  
Keyword(s):  
Membrane Damage ◽  
Coiled Coil ◽  
Cardiac Cells ◽  
Acute Injury ◽  
Membrane Repair ◽  
Repair Capacity ◽  
E3 Ligases ◽  
Tripartite Motif ◽  
Trim Proteins

Tripartite motif (TRIM) proteins are a superfamily of coiled-coil-containing RING E3 ligases that function in many cellular processes, particularly in membrane repair pathways. Mitsugumin 53 (MG53) also known as TRIM72, is primary expressed in skeletal muscle and heart. Our experimental data confirm that during membrane damage, MG53 translocates to the injury site and acts as a molecular glue to reseal the damage area. The role of MG53 in membrane repair has been demonstrated in both in vitro studies using molecular approaches and in vivo using rodent wild type and knockout models. Thus, our data indicate that recombinant human MG53 protein can be directly applied as a therapeutic agent to increase the membrane repair capacity of many cell types, including cardiomyocytes during acute injury or in chronic disease progression. However, the precise mechanism and potential partners by which MG53 executes its membrane repair function are not completely understood. On the basis of the global TRIM family protein alignment, we hypothesize that there are other TRIM proteins that, alone or together with MG53, may facilitate repair by targeting the site of an injury. Moreover, data from our lab demonstrated that MG53 and these TRIM proteins can form homo- and hetero-oligomeric assemblies due to the presence of the coiled-coil region in these proteins and, further, that this may be necessary for the active membrane resealing process. Using E. coli protein expression methodology we can generate and isolate new TRIM recombinant proteins and test if these protein complexes are effective when applied externally to cardiac and non-cardiac cells. These novel proteins will also be tested for their pharmacokinetic properties to determine their efficacy in both acute and chronic applications. Our studies should increase our knowledge of the mechanisms controlling cardiac membrane repair and also provide novel therapeutic targets.

scholarly journals SVIP Is a Novel VCP/p97-interacting Protein Whose Expression Causes Cell Vacuolation

2003 ◽  
Vol 14 (1) ◽  
pp. 262-273 ◽  
Author(s):  
Masami Nagahama ◽  
Mie Suzuki ◽  
Yuko Hamada ◽  
Kiyotaka Hatsuzawa ◽  
Katsuko Tani ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Coiled Coil ◽  
Adaptor Proteins ◽  
Interacting Protein ◽  
Cellular Processes ◽  
Coiled Coil Region ◽  
Acid Protein ◽  
Extensive Cell ◽  
Dependent Protein ◽  
Amino Acid Protein

VCP/p97 is involved in a variety of cellular processes, including membrane fusion and ubiquitin-dependent protein degradation. It has been suggested that adaptor proteins such as p47 and Ufd1p confer functional versatility to VCP/p97. To identify novel adaptors, we searched for proteins that interact specifically with VCP/p97 by using the yeast two-hybrid system, and discovered a novel VCP/p97-interacting protein named smallVCP/p97-interactingprotein (SVIP). Rat SVIP is a 76-amino acid protein that contains two putative coiled-coil regions, and potential myristoylation and palmitoylation sites at the N terminus. Binding experiments revealed that the N-terminal coiled-coil region of SVIP, and the N-terminal and subsequent ATP-binding regions (ND1 domain) of VCP/p97, interact with each other. SVIP and previously identified adaptors p47 and ufd1p interact with VCP/p97 in a mutually exclusive manner. Overexpression of full-length SVIP or a truncated mutant did not markedly affect the structure of the Golgi apparatus, but caused extensive cell vacuolation reminiscent of that seen upon the expression of VCP/p97 mutants or polyglutamine proteins in neuronal cells. The vacuoles seemed to be derived from endoplasmic reticulum membranes. These results together suggest that SVIP is a novelVCP/p97 adaptor whose function is related to the integrity of the endoplasmic reticulum.

scholarly journals Does it take two to tango? RING domain self-association and activity in TRIM E3 ubiquitin ligases

2020 ◽  
Vol 48 (6) ◽  
pp. 2615-2624
Author(s):  
Filippo Fiorentini ◽  
Diego Esposito ◽  
Katrin Rittinger
Keyword(s):  
Coiled Coil ◽  
E3 Ligase ◽  
Protein Family ◽  
Ring Domain ◽  
Cellular Functions ◽  
Protein Activity ◽  
Coiled Coil Region ◽  
Tripartite Motif ◽  
Large Protein Family ◽  
Self Association

TRIM proteins form a protein family that is characterized by a conserved tripartite motif domain comprising a RING domain, one or two B-box domains and a coiled-coil region. Members of this large protein family are important regulators of numerous cellular functions including innate immune responses, transcriptional regulation and apoptosis. Key to their cellular role is their E3 ligase activity which is conferred by the RING domain. Self-association is an important characteristic of TRIM protein activity and is mediated by homodimerization via the coiled-coil region, and in some cases higher order association via additional domains of the tripartite motif. In many of the TRIM family proteins studied thus far, RING dimerization is an important prerequisite for E3 ligase enzymatic activity though the propensity of RING domains to dimerize differs significantly between different TRIMs and can be influenced by other regions of the protein.

scholarly journals TRIMming down to TRIM37: Relevance to Inflammation, Cardiovascular Disorders, and Cancer in MULIBREY Nanism

2018 ◽  
Vol 20 (1) ◽  
pp. 67 ◽  
Author(s):  
Benjamin Brigant ◽  
Valérie Metzinger-Le Meuth ◽  
Jacques Rochette ◽  
Laurent Metzinger
Keyword(s):  
Genetic Disorder ◽  
Coiled Coil ◽  
Growth Failure ◽  
Postnatal Growth ◽  
E3 Ligases ◽  
Domain Specific ◽  
Mulibrey Nanism ◽  
Glucose And Lipid Metabolism ◽  
Coiled Coil Region ◽  
Innovative Drugs

TRIpartite motif (TRIM) proteins are part of the largest subfamilies of E3 ligases that mediate the transfer of ubiquitin to substrate target proteins. In this review, we focus on TRIM37 in the normal cell and in pathological conditions, with an emphasis on the MULIBREY (MUscle-LIver-BRain-EYe) genetic disorder caused by TRIM37 mutations. TRIM37 is characterized by the presence of a RING domain, B-box motifs, and a coiled-coil region, and its C-terminal part includes the MATH domain specific to TRIM37. MULIBREY nanism is a rare autosomal recessive caused by TRIM37 mutations and characterized by severe pre- and postnatal growth failure. Constrictive pericarditis is the most serious anomaly of the disease and is present in about 20% of patients. The patients have a deregulation of glucose and lipid metabolism, including type 2 diabetes, fatty liver, and hypertension. Puzzlingly, MULIBREY patients, deficient for TRIM37, are plagued with numerous tumors. Among non-MULIBREY patients affected by cancer, a wide variety of cancers are associated with an overexpression of TRIM37. This suggests that normal cells need an optimal equilibrium in TRIM37 expression. Finding a way to keep that balance could lead to potential innovative drugs for MULIBREY nanism, including heart condition and carcinogenesis treatment.

scholarly journals A helical LC3-interacting region mediates the interaction between the retroviral restriction factor Trim5α and mammalian autophagy-related ATG8 proteins

2018 ◽  
Vol 293 (47) ◽  
pp. 18378-18386 ◽  
Author(s):  
Jeremy R. Keown ◽  
Moyra M. Black ◽  
Aaron Ferron ◽  
Melvyn Yap ◽  
Michael J. Barnett ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Life Cycle ◽  
Analytical Ultracentrifugation ◽  
Coiled Coil ◽  
Restriction Factor ◽  
Related Protein ◽  
Binding Motifs ◽  
Binding Interface ◽  
Coiled Coil Region ◽  
Tripartite Motif

The retroviral restriction factor tripartite motif–containing 5α (Trim5α) acts during the early postentry stages of the retroviral life cycle to block infection by a broad range of retroviruses, disrupting reverse transcription and integration. The mechanism of this restriction is poorly understood, but it has recently been suggested to involve recruitment of components of the autophagy machinery, including members of the mammalian autophagy-related 8 (ATG8) family involved in targeting proteins to the autophagosome. To better understand the molecular details of this interaction, here we utilized analytical ultracentrifugation to characterize the binding of six ATG8 isoforms and determined the crystal structure of the Trim5α Bbox coiled-coil region in complex with one member of the mammalian ATG8 proteins, autophagy-related protein LC3 B (LC3B). We found that Trim5α binds all mammalian ATG8s and that, unlike the typical LC3-interacting region (LIR) that binds to mammalian ATG8s through a β-strand motif comprising approximately six residues, LC3B binds to Trim5α via the α-helical coiled-coil region. The orientation of the structure demonstrated that LC3B could be accommodated within a Trim5α assembly that can bind the retroviral capsid. However, mutation of the binding interface does not affect retroviral restriction. Comparison of the typical linear β-strand LIR with our atypical helical LIR reveals a conservation of the presentation of residues that are required for the interaction with LC3B. This observation expands the range of LC3B-binding proteins to include helical binding motifs and demonstrates a link between Trim5α and components of the autophagosome.

scholarly journals Structural insights into WHAMM-mediated cytoskeletal coordination during membrane remodeling

2012 ◽  
Vol 199 (1) ◽  
pp. 111-124 ◽  
Author(s):  
Qing-Tao Shen ◽  
Peter P. Hsiue ◽  
Charles V. Sindelar ◽  
Matthew D. Welch ◽  
Kenneth G. Campellone ◽  
...  
Keyword(s):  
Intracellular Transport ◽  
Coiled Coil ◽  
Tubular Structures ◽  
Terminal Portion ◽  
Actin Nucleation ◽  
Cellular Processes ◽  
Coiled Coil Region ◽  
Membrane Tubulation ◽  
Underlying Mechanisms ◽  
Structural Insights

The microtubule (MT) and actin cytoskeletons drive many essential cellular processes, yet fairly little is known about how their functions are coordinated. One factor that mediates important cross talk between these two systems is WHAMM, a Golgi-associated protein that utilizes MT binding and actin nucleation activities to promote membrane tubulation during intracellular transport. Using cryoelectron microscopy and other biophysical and biochemical approaches, we unveil the underlying mechanisms for how these activities are coordinated. We find that WHAMM bound to the outer surface of MT protofilaments via a novel interaction between its central coiled-coil region and tubulin heterodimers. Upon the assembly of WHAMM onto MTs, its N-terminal membrane-binding domain was exposed at the MT periphery, where it can recruit vesicles and remodel them into tubular structures. In contrast, MT binding masked the C-terminal portion of WHAMM and prevented it from promoting actin nucleation. These results give rise to a model whereby distinct MT-bound and actin-nucleating populations of WHAMM collaborate during membrane tubulation.

scholarly journals Functional Replacement of the RING, B-Box 2, and Coiled-Coil Domains of Tripartite Motif 5α (TRIM5α) by Heterologous TRIM Domains

2006 ◽  
Vol 80 (13) ◽  
pp. 6198-6206 ◽  
Author(s):  
Xing Li ◽  
Yuan Li ◽  
Matthew Stremlau ◽  
Wen Yuan ◽  
Byeongwoon Song ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Coiled Coil ◽  
Chimeric Proteins ◽  
Immunodeficiency Virus ◽  
Tripartite Motif ◽  
Trim Proteins ◽  
Hiv 1 ◽  
Functional Replacement ◽  
Distant Relative

ABSTRACT Tripartite motif 5α (TRIM5α) restricts some retroviruses, including human immunodeficiency virus type 1 (HIV-1), from infecting the cells of particular species. TRIM5α is a member of the TRIM family of proteins, which contain RING, B-box, coiled-coil (CC), and, in some cases, B30.2(SPRY) domains. Here we investigated the abilities of domains from TRIM proteins (TRIM6, TRIM34, and TRIM21) that do not restrict HIV-1 infection to substitute for the domains of rhesus monkey TRIM5α (TRIM5αrh). The RING, B-box 2, and CC domains of the paralogous TRIM6 and TRIM34 proteins functionally replaced the corresponding TRIM5αrh domains, allowing HIV-1 restriction. By contrast, similar chimeras containing the components of TRIM21, a slightly more distant relative of TRIM5, did not restrict HIV-1 infection. The TRIM21 B-box 2 domain and its flanking linker regions contributed to the functional defectiveness of these chimeras. All of the chimeric proteins formed trimers. All of the chimeras that restricted HIV-1 infection bound the assembled HIV-1 capsid complexes. These results indicate that heterologous RING, B-box 2, and CC domains from related TRIM proteins can functionally substitute for TRIM5αrh domains.

scholarly journals Structural analysis of RIG-I-like receptors reveals ancient rules of engagement between diverse RNA helicases and TRIM ubiquitin ligases

2020 ◽  
Author(s):  
Kazuki Kato ◽  
Sadeem Ahmad ◽  
Zixiang Zhu ◽  
Janet M. Young ◽  
Xin Mu ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Rna Helicases ◽  
Animal Evolution ◽  
E3 Ligases ◽  
Cryo Electron Microscopy ◽  
Rna Biology ◽  
Evolutionarily Conserved ◽  
Tripartite Motif ◽  
Biochemical Analyses ◽  
Trim Proteins

AbstractRNA helicases and ubiquitin E3 ligases mediate many critical functions within cells, but their actions have been studied largely in distinct biological contexts. Here, we uncover evolutionarily conserved rules of engagement between RNA helicases and tripartite motif (TRIM) E3 ligases that lead to their functional coordination in vertebrate innate immunity. Using cryo-electron microscopy and biochemistry, we show that RIG-I-like receptors (RLRs), viral RNA receptors with helicase domains, interact with their cognate TRIM/TRIM-like E3 ligases through similar epitopes in the helicase domains. Their interactions are avidity-driven, restricting the actions of TRIM/TRIM-like proteins and consequent immune activation to RLR multimers. Mass-spectrometry and phylogeny-guided biochemical analyses further reveal that similar rules of engagement apply to diverse RNA helicases and TRIM/TRIM-like proteins. Our analyses thus reveal not only conserved substrates for TRIM proteins but also unexpectedly deep evolutionary connections between TRIM proteins and RNA helicases, thereby linking ubiquitin and RNA biology throughout animal evolution.

scholarly journals TRIM protein domain topology and implications for antiviral immunity

2014 ◽  
Vol 70 (a1) ◽  
pp. C243-C243
Author(s):  
David Jacques ◽  
Cy Jeffries ◽  
Matthew Caines ◽  
Michael Lammers ◽  
Donna Mallery ◽  
...  
Keyword(s):  
Domain Architecture ◽  
Coiled Coil ◽  
Antiviral Immunity ◽  
Protein Domain ◽  
Titration Calorimetry ◽  
Protein Chain ◽  
Trim Protein ◽  
Tripartite Motif ◽  
Trim Proteins ◽  
Terminal Domains

The tripartite motif (TRIM) proteins are a large family of >100 members, several of which have important roles in antiviral immunity and innate immune signaling. TRIM5α associates with incoming HIV-1 capsids, interfering with controlled disassembly and targeting them for degradation by the proteasome. TRIM21 is a cytosolic antibody receptor, which also targets incoming viral capsids for proteasomal degradation. TRIM25 is also involved in innate immunity, being essential for the ubiquitination of RIG-I. Recent positive selection analysis has predicted another 10 TRIM proteins with antiviral activity. Despite the fact that TRIM5α, 21 and 25 play key roles in antiviral protection, their mechanism of action is incompletely understood. All three proteins share a similar domain architecture, comprising a RING, B Box, coiled coil and PRYSPRY domains. The RING domains are responsible for ubiquitin ligase activity, while the PRYSPRY domains determine target specificity. We have used a combination of crystallography and SAXS to generate the first complete model for a TRIM protein structure. Crystallographic studies of TRIM25 reveal a central elongated coiled-coil domain with an unusual right-handed twist. The dimer formed by the coiled-coil is antiparallel but is followed by additional helices that reverse the direction of the protein chain. This structure suggests that the N-terminal domains of each monomer are separated but the C terminal domains are maintained in proximity. Multi-angle light scattering (MALS), isothermal titration calorimetry (ITC) and SAXS analysis confirms that this dimer structure is present in solution. Furthermore, scattering studies on the tripartite motif of TRIM21, comprising RING, B Box and coiled-coil, demonstrate that the first two domains of each monomer are held 150-200 Å apart. Finally, SAXS measurement of a complex between intact TRIM21 and its ligand, IgG Fc, provides the first empirical structure of a complete TRIM protein.

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