Structure of the rhesus monkey TRIM5α PRYSPRY domain, the HIV capsid recognition module

Biris, Nikolaos; Yang, Yang; Taylor, Alexander B.; Tomashevski, Andrei; Guo, Miao; Hart, P. John; Díaz-Griffero, Felipe; Ivanov, Dmitri N.

doi:10.1073/pnas.1203536109

Cited by 82 publications

(103 citation statements)

References 49 publications

(63 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A similar conclusion has been reached from studies of different lentiviruses (38)(39)(40)(41)(42). Such an extended interaction domain would be consistent with structural studies of the TRIM5␣ PRY-SPRY region that suggest a large, triangular surface region containing the specificity determinants of restriction (25,43,44). Studies of CA determinants of Fv1 tropism also suggest an extended interaction domain (45).…”

Section: Discussionsupporting

confidence: 66%

A Comparison of Murine Leukemia Viruses That Escape from Human and Rhesus Macaque TRIM5αs

Ohkura¹,

Stoye²

2013

J Virol

View full text Add to dashboard Cite

To better understand the binding mechanism of TRIM5␣ to retrovirus capsid, we had previously selected N-tropic murine leukemia virus (N-MLV) mutants escaping from rhesus macaque TRIM5␣ (rhTRIM5␣) by passaging the virus in rhTRIM5␣-expressing cells and selecting for nonrestricted variants. To test the commonality of the findings from the rhTRIM5␣ study, we have now employed a similar genetic approach using human TRIM5␣ (huTRIM5␣). Consistent with the rhTRIM5␣ study, the mapped huTRIM5␣ escape mutations were distributed across the capsid exterior, confirming the extended binding surface between virus and restriction factor. Compared to the results of the previous study, fewer escape mutations were identified, with particular mutants being repeatedly selected. Three out four huTRIM5␣ escape variants showed resistance to all primate TRIM5␣s tested, but two of them sacrificed viral fitness, observations that were not made in the rhTRIM5␣ study. Moreover, differences in amino acid changes associated with escape from hu-and rhTRIM5␣s suggested a charge dependence of the restriction by different TRIM5␣s. Taken together, these results suggest that the recognition of the entire capsid surface is a general strategy for TRIM5␣ to restrict MLV but that significantly different specific interactions are involved in the binding of TRIM5␣ from different species to the MLV capsid core. C apsid (CA)-binding retrovirus restriction factors have arisen on at least five occasions in the course of evolution. Their independent evolution is testament to the utility of such factors, either to limit endogenous retrovirus amplification or to inhibit infection by exogenous viruses (1-3). This group of factors includes Fv1 (4, 5), TRIM5␣ (6), and three independently arising hybrid factors, TRIM5CypA1 (7, 8), TRIM5CypA2 (9-12), and TRIM5CypA3 (13) in which a retrotransposed copy of the cellular cyclophilin-A gene has been inserted in the 3= end of the TRIM5 gene. At a minimum, these factors contain a virus CA-binding domain, as well as one or more multimerization domains (14-22).Restriction factor engagement appears to occur soon after retroviral core release into the cell cytoplasm, resulting either in proteasome-mediated CA degradation and an inhibition of reverse transcription (6) or an apparent sequestration of the reverse-transcribed preintegration complex en route to the cell nucleus (23).One of the least-appreciated aspects of restriction by these factors is the ability to restrict multiple species of retrovirus, a property that presumably reflects the ability of these factors to bind several different CA molecules. For example, TRIM5␣ from the cotton-top tamarin can restrict one or more members of the lentivirus, gammaretrovirus, betaretrovirus, and foamy virus genera (24-26), despite extensive differences in the sequences of their CA molecules. Nevertheless, restriction can be sensitive to a single amino acid change, as best characterized by the single change at CA position 110 of murine leukemia virus (MLV) that distinguishes between...

show abstract

Section: Discussionsupporting

confidence: 66%

A Comparison of Murine Leukemia Viruses That Escape from Human and Rhesus Macaque TRIM5αs

Ohkura¹,

Stoye²

2013

J Virol

View full text Add to dashboard Cite

show abstract

“…The 4 hyper-variable regions of TRIM5α are present at the surface of the protein and constitute the binding interface for interactions with retroviral capsids, as has been confirmed recently (Biris et al, 2012). We used an in silico assay to predict the impact of mutations at positions 330, 332 and 335 on this CA-binding interface (Fig.…”

Section: Resultssupporting

confidence: 54%

“…The I-TASSER program integrated more than 10 published structures of domains similar to the WT TRIM5α Hu PRYSPRY domain. Among those was the recently published structure of the Rhesus macaque TRIM5α PRYSPRY (Biris et al, 2012). According to this prediction tool, introduction of the R332G and R335G mutations caused significant changes in all 3 main variable regions (v1, v2, v3).…”

Section: Resultsmentioning

confidence: 99%

A novel aminoacid determinant of HIV-1 restriction in the TRIM5α variable 1 region isolated in a random mutagenic screen

et al. 2013

Self Cite

View full text Add to dashboard Cite

Human-derived antiretroviral transgenes are of great biomedical interest and are actively pursued. HIV-1 is efficiently inhibited at post-entry, pre-integration replication stages by point mutations in the variable region 1 (v1) of the human restriction factor TRIM5α. Here we use a mutated megaprimer approach to create a mutant library of TRIM5αHu v1 and to isolate a mutation at Gly330 (G330E) that inhibits transduction of an HIV-1 vector as efficiently as the previously described mutants at positions Arg332 and Arg335. As was the case for these other mutations, modification of the local v1 charge toward increased acidity was key to inhibiting HIV-1. G330E TRIM5αHu also disrupted replication-competent HIV-1 propagation in a human T cell line. Interestingly, G330E did not enhance restriction of HIV-1 when combined with mutations at Arg332 or Arg335. Accordingly, the triple mutant G330E-R332G–R335G bound purified recombinant HIV-1 capsid tubes less efficiently than the double mutant R332G–R335G did. In a structural model of the TRIM5αHu PRYSPRY domain, the addition of G330E to the double mutant R332G–R335G caused extensive changes to the capsid-binding surface, which may explain why the triple mutant was no more restrictive than the double mutant. The HIV-1 inhibitory potential of Gly330 mutants was not predicted by examination of natural TRIM5α orthologs that are known to strongly inhibit HIV-1. This work underlines the potential of random mutagenesis to isolate novel variants of human proteins with antiviral properties.

show abstract

“…Interestingly, residues 291-300 of rhesus TRIM5α (colored in blue and boxed in Fig. S7) do indeed form a helix in two independent crystal structures of the isolated B30.2 domain (42,43), although the immediately preceding residues (287-290) adopt a nonhelical loop configuration with high temperature factors in one of the structures (43). These observations lead us to speculate that the N-terminal helix of the TRIM5α B30.2 domain (or the longer, predicted helix) may pack against the center of the upstream coiled-coil to form a 4-helix bundle, as seen in the TRIM25 structure.…”

Section: Discussionmentioning

confidence: 99%

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

Sanchez

Okreglicka

Chandrasekaran

et al. 2014

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

160

218

View full text Add to dashboard Cite

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...

show abstract

Structure of the rhesus monkey TRIM5α PRYSPRY domain, the HIV capsid recognition module

Cited by 82 publications

References 49 publications

A Comparison of Murine Leukemia Viruses That Escape from Human and Rhesus Macaque TRIM5αs

A Comparison of Murine Leukemia Viruses That Escape from Human and Rhesus Macaque TRIM5αs

A novel aminoacid determinant of HIV-1 restriction in the TRIM5α variable 1 region isolated in a random mutagenic screen

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

Contact Info

Product

Resources

About