Determinants of the Higher Order Association of the Restriction Factor TRIM5α and Other Tripartite Motif (TRIM) Proteins

Li, Xing; Yeung, Darwin F.; Fiegen, Ann M.; Sodroski, Joseph

doi:10.1074/jbc.m111.260406

Cited by 61 publications

(71 citation statements)

References 46 publications

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“…In the alternative configuration, the associated coiled-coil dimers would "swap" arms in a fashion reminiscent of clathrin triskelion assembly (39). We cannot yet unambiguously discriminate between these different possible assembly modes but note that domain swapping would provide a mechanism for autoinhibition, and thereby prevent unregulated assembly, and would be consistent with recent studies indicating that the L2 linker plays an important role in high-order TRIM5α assembly (40,41).…”

Section: Discussionsupporting

confidence: 62%

The tripartite motif coiled-coil is an elongated antiparallel hairpin dimer

Sanchez

Okreglicka

Chandrasekaran

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

171

231

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Tripartite motif (TRIM) proteins make up a large family of coiledcoil-containing RING E3 ligases that function in many cellular processes, particularly innate antiviral response pathways. Both dimerization and higher-order assembly are important elements of TRIM protein function, but the atomic details of TRIM tertiary and quaternary structure have not been fully understood. Here, we present crystallographic and biochemical analyses of the TRIM coiled-coil and show that TRIM proteins dimerize by forming interdigitating antiparallel helical hairpins that position the Nterminal catalytic RING domains at opposite ends of the dimer and the C-terminal substrate-binding domains at the center. The dimer core comprises an antiparallel coiled-coil with a distinctive, symmetric pattern of flanking heptad and central hendecad repeats that appear to be conserved across the entire TRIM family. Our studies reveal how the coiled-coil organizes TRIM25 to polyubiquitylate the RIG-I/viral RNA recognition complex and how dimers of the TRIM5α protein are arranged within hexagonal arrays that recognize the HIV-1 capsid lattice and restrict retroviral replication.antiparallel dimer | disulfide crosslinking | X-ray crystallography T ripartite motif (TRIM) proteins make up the largest superfamily of RING E3 ubiquitin ligases, with more than 100 members in the human proteome (1, 2). TRIM proteins function in a variety of cellular pathways, and many regulate innate immunity and/or mediate antiviral responses. Antiviral TRIMs include TRIM25, which regulates the IFN response to RNA viruses (3, 4), and TRIM5α, which senses and inhibits early stages of retroviral replication (5, 6).TRIM proteins share a common N-terminal domain organization, termed the tripartite or RBCC (RING, B-box, coiled-coil) motif, followed by variable C-terminal protein recognition domains ( Fig. 1 A and B). "Linker" segments of unknown structure typically separate both the RING and B-box domains (L1) and the coiledcoil and terminal effector domains (L2). The coiled-coil region mediates oligomerization, and both homooligomeric and heterooligomeric TRIMs have been described (7-13). Furthermore, many TRIM proteins form higher-order assemblies in vitro and form punctate or fibrous structures in cells (14-16). For example, TRIM5α assembly allows the protein to function as a cytosolic patternrecognition receptor that can intercept the incoming capsids of diverse retroviruses, including HIV-1 (6). This results in species-specific "restriction" of viral replication (5), capsid dissociation (5, 6), and induction of innate immune responses (17). Retroviral capsids are recognized through a remarkable mechanism of multivalent pattern recognition. TRIM5α forms a homodimer (10, 11, 18), which can further assemble into a 2D lattice of linked hexagons (18). The hexagonal TRIM5α net matches the symmetry and spacing of the retroviral capsid surface lattice, thereby positioning multiple C-terminal B30.2/SPRY domains to interact with their repeating binding epitopes on the capsid.Struct...

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Section: Discussionsupporting

confidence: 62%

The tripartite motif coiled-coil is an elongated antiparallel hairpin dimer

Sanchez

Okreglicka

Chandrasekaran

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

171

231

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“…The B‐boxes of TRIM25 and TRIM32 have no apparent effect on oligomerization and only a small effect on catalytic activity, clearly setting them apart from the best studied member of the TRIM family, the retroviral restriction factor TRIM5α, in which the B‐box promotes higher order oligomerization (Diaz‐Griffero et al , 2009; Li et al , 2011). This raises questions about the role of B‐boxes in those TRIMs that do not require additional oligomerization domains.…”

Section: Discussionmentioning

confidence: 99%

“…All TRIM ligases contain at least B‐box2 while those with tandem B‐boxes contain one of each with B‐box1 located N‐terminal to B‐box2. B‐box domains act as protein–protein interaction motifs and mediate self‐association in some TRIMs, which may be important for higher order TRIM oligomerization as seen in the case of TRIM5α (Tao et al , 2008; Diaz‐Griffero et al , 2009; Li et al , 2011). …”

Section: Introductionmentioning

confidence: 99%

“…Such an anti‐parallel arrangement locates the N‐terminal RING domains at opposite ends of the molecule implying that a dimeric RING arrangement may require formation of higher order TRIM complexes. For some TRIMs such as the retroviral restriction factor TRIM5α, higher order complexes have been shown to exist and constitute the active form (Diaz‐Griffero et al , 2009; Li et al , 2011), whereas others, such as TRIM25, have been shown to be solely dimeric (Li et al , 2014). …”

Section: Introductionmentioning

confidence: 99%

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Functional role of TRIM E3 ligase oligomerization and regulation of catalytic activity

et al. 2016

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TRIM E3 ubiquitin ligases regulate a wide variety of cellular processes and are particularly important during innate immune signalling events. They are characterized by a conserved tripartite motif in their N‐terminal portion which comprises a canonical RING domain, one or two B‐box domains and a coiled‐coil region that mediates ligase dimerization. Self‐association via the coiled‐coil has been suggested to be crucial for catalytic activity of TRIMs; however, the precise molecular mechanism underlying this observation remains elusive. Here, we provide a detailed characterization of the TRIM ligases TRIM25 and TRIM32 and show how their oligomeric state is linked to catalytic activity. The crystal structure of a complex between the TRIM25 RING domain and an ubiquitin‐loaded E2 identifies the structural and mechanistic features that promote a closed E2~Ub conformation to activate the thioester for ubiquitin transfer allowing us to propose a model for the regulation of activity in the full‐length protein. Our data reveal an unexpected diversity in the self‐association mechanism of TRIMs that might be crucial for their biological function.

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“…PRY/SPRY rh to the C-terminal helix of MBP. This segment is part of the L2 region (residues 234-296) of TRIM5α (28) and is presumably unstructured because the majority of the L2 residues are not included in the construct.…”

Section: Resultsmentioning

confidence: 99%

Structural insight into HIV-1 capsid recognition by rhesus TRIM5α

Yang

Zhao

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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Tripartite motif protein isoform 5 alpha (TRIM5α) is a potent antiviral protein that restricts infection by HIV-1 and other retroviruses. TRIM5α recognizes the lattice of the retrovirus capsid through its B30.2 (PRY/SPRY) domain in a species-specific manner. Upon binding, TRIM5α induces premature disassembly of the viral capsid and activates the downstream innate immune response. We have determined the crystal structure of the rhesus TRIM5α PRY/ SPRY domain that reveals essential features for capsid binding. Combined cryo-electron microscopy and biochemical data show that the monomeric rhesus TRIM5α PRY/SPRY, but not the human TRIM5α PRY/SPRY, can bind to HIV-1 capsid protein assemblies without causing disruption of the capsid. This suggests that the PRY/SPRY domain alone constitutes an important pattern-sensing component of TRIM5α that is capable of interacting with viral capsids of different curvatures. Our results provide molecular insights into the mechanisms of TRIM5α-mediated retroviral restriction.

show abstract

Determinants of the Higher Order Association of the Restriction Factor TRIM5α and Other Tripartite Motif (TRIM) Proteins

Cited by 61 publications

References 46 publications

The tripartite motif coiled-coil is an elongated antiparallel hairpin dimer

The tripartite motif coiled-coil is an elongated antiparallel hairpin dimer

Functional role of TRIM E3 ligase oligomerization and regulation of catalytic activity

Structural insight into HIV-1 capsid recognition by rhesus TRIM5α

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