SUMO-Interacting Motifs of Human TRIM5α are Important for
Antiviral Activity


Human TRIM5α potently restricts particular strains of murine leukemia viruses

(the so-called N-tropic strains) but not others (the B- or NB-tropic strains)

during early stages of infection. We show that overexpression of SUMO-1 in human

293T cells, but not in mouse MDTF cells, profoundly blocks N-MLV infection. This

block is dependent on the tropism of the incoming virus, as neither B-, NB-, nor

the mutant R110E of N-MLV CA (a B-tropic switch) are affected by SUMO-1

overexpression. The block occurred prior to reverse transcription and could be

abrogated by large amounts of restricted virus. Knockdown of TRIM5α in 293T

SUMO-1-overexpressing cells resulted in ablation of the SUMO-1 antiviral

effects, and this loss of restriction could be restored by expression of a human

TRIM5α shRNA-resistant plasmid. Amino acid sequence analysis of human

TRIM5α revealed a consensus SUMO conjugation site at the N-terminus and

three putative SUMO interacting motifs (SIMs) in the B30.2 domain. Mutations of

the TRIM5α consensus SUMO conjugation site did not affect the antiviral

activity of TRIM5α in any of the cell types tested. Mutation of the SIM

consensus sequences, however, abolished TRIM5α antiviral activity against

N-MLV. Mutation of lysines at a potential site of SUMOylation in the CA region

of the Gag gene reduced the SUMO-1 block and the TRIM5α restriction of

N-MLV. Our data suggest a novel aspect of TRIM5α-mediated restriction, in

which the presence of intact SIMs in TRIM5α, and also the SUMO conjugation

of CA, are required for restriction. We propose that at least a portion of the

antiviral activity of TRIM5α is mediated through the binding of its SIMs to

SUMO-conjugated CA.


Vyšlo v časopise: SUMO-Interacting Motifs of Human TRIM5α are Important for Antiviral Activity. PLoS Pathog 7(4): e32767. doi:10.1371/journal.ppat.1002019
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1002019

Souhrn

Human TRIM5α potently restricts particular strains of murine leukemia viruses

(the so-called N-tropic strains) but not others (the B- or NB-tropic strains)

during early stages of infection. We show that overexpression of SUMO-1 in human

293T cells, but not in mouse MDTF cells, profoundly blocks N-MLV infection. This

block is dependent on the tropism of the incoming virus, as neither B-, NB-, nor

the mutant R110E of N-MLV CA (a B-tropic switch) are affected by SUMO-1

overexpression. The block occurred prior to reverse transcription and could be

abrogated by large amounts of restricted virus. Knockdown of TRIM5α in 293T

SUMO-1-overexpressing cells resulted in ablation of the SUMO-1 antiviral

effects, and this loss of restriction could be restored by expression of a human

TRIM5α shRNA-resistant plasmid. Amino acid sequence analysis of human

TRIM5α revealed a consensus SUMO conjugation site at the N-terminus and

three putative SUMO interacting motifs (SIMs) in the B30.2 domain. Mutations of

the TRIM5α consensus SUMO conjugation site did not affect the antiviral

activity of TRIM5α in any of the cell types tested. Mutation of the SIM

consensus sequences, however, abolished TRIM5α antiviral activity against

N-MLV. Mutation of lysines at a potential site of SUMOylation in the CA region

of the Gag gene reduced the SUMO-1 block and the TRIM5α restriction of

N-MLV. Our data suggest a novel aspect of TRIM5α-mediated restriction, in

which the presence of intact SIMs in TRIM5α, and also the SUMO conjugation

of CA, are required for restriction. We propose that at least a portion of the

antiviral activity of TRIM5α is mediated through the binding of its SIMs to

SUMO-conjugated CA.


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