Endoplasmic reticulum chaperones stabilize ligand-receptive MR1 molecules for efficient presentation of metabolite antigens

Hamish E. G. McWilliam; Jeffrey Y. W. Mak; Wael Awad; Matthew  Zorkau; Sebastian Cruz-Gomez; Hui Jing Lim; Yuting Yan; Sam Wormald; Laura F. Dagley; Sidonia B. G. Eckle; Alexandra J. Corbett; Haiyin Liu; Shihan Li; Scott J. J. Reddiex; Justine D. Mintern; Ligong Liu; James McCluskey; Jamie Rossjohn; David P. Fairlie; Jose A. Villadangos

doi:10.1073/pnas.2011260117

Endoplasmic reticulum chaperones stabilize ligand-receptive MR1 molecules for efficient presentation of metabolite antigens

Hamish E. G. McWilliam, Jeffrey Y. W. Mak, Wael Awad, Matthew Zorkau, Sebastian Cruz-Gomez, Hui Jing Lim, Yuting Yan, Sam Wormald, Laura F. Dagley, Sidonia B. G. Eckle, Alexandra J. Corbett, Haiyin Liu, Shihan Li, Scott J. J. Reddiex, Justine D. Mintern, Ligong Liu, James McCluskey, Jamie Rossjohn, David P. Fairlie, Jose A. Villadangos

Research output: Contribution to journal › Article › Research › peer-review

38 Citations (Scopus)

Abstract

The antigen-presenting molecule MR1 (MHC class I-related protein 1) presents metabolite antigens derived from microbial vitamin B2 synthesis to activate mucosal-associated invariant T (MAIT) cells. Key aspects of this evolutionarily conserved pathway remain uncharacterized, including where MR1 acquires ligands and what accessory proteins assist ligand binding. We answer these questions by using a fluorophore-labeled stable MR1 antigen analog, a conformation-specific MR1 mAb, proteomic analysis, and a genome-wide CRISPR/Cas9 library screen. We show that the endoplasmic reticulum (ER) contains a pool of two unliganded MR1 conformers stabilized via interactions with chaperones tapasin and tapasin-related protein. This pool is the primary source of MR1 molecules for the presentation of exogenous metabolite antigens to MAIT cells. Deletion of these chaperones reduces the ERresident MR1 pool and hampers antigen presentation and MAIT cell activation. The MR1 antigen-presentation pathway thus co-opts ER chaperones to fulfill its unique ability to present exogenous metabolite antigens captured within the ER.

Original language	English
Pages (from-to)	24974-24985
Number of pages	12
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	117
Issue number	40
DOIs	https://doi.org/10.1073/pnas.2011260117
Publication status	Published - 6 Oct 2020

Keywords

Fluorescent probe
MAIT cells
MHC class I-related protein 1 (MR1)
Protein trafficking
Vitamin B

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10.1073/pnas.2011260117

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McWilliam, H. E. G., Mak, J. Y. W., Awad, W., Zorkau, M., Cruz-Gomez, S., Lim, H. J., Yan, Y., Wormald, S., Dagley, L. F., Eckle, S. B. G., Corbett, A. J., Liu, H., Li, S., Reddiex, S. J. J., Mintern, J. D., Liu, L., McCluskey, J., Rossjohn, J., Fairlie, D. P., & Villadangos, J. A. (2020). Endoplasmic reticulum chaperones stabilize ligand-receptive MR1 molecules for efficient presentation of metabolite antigens. Proceedings of the National Academy of Sciences of the United States of America, 117(40), 24974-24985. https://doi.org/10.1073/pnas.2011260117

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title = "Endoplasmic reticulum chaperones stabilize ligand-receptive MR1 molecules for efficient presentation of metabolite antigens",

abstract = "The antigen-presenting molecule MR1 (MHC class I-related protein 1) presents metabolite antigens derived from microbial vitamin B2 synthesis to activate mucosal-associated invariant T (MAIT) cells. Key aspects of this evolutionarily conserved pathway remain uncharacterized, including where MR1 acquires ligands and what accessory proteins assist ligand binding. We answer these questions by using a fluorophore-labeled stable MR1 antigen analog, a conformation-specific MR1 mAb, proteomic analysis, and a genome-wide CRISPR/Cas9 library screen. We show that the endoplasmic reticulum (ER) contains a pool of two unliganded MR1 conformers stabilized via interactions with chaperones tapasin and tapasin-related protein. This pool is the primary source of MR1 molecules for the presentation of exogenous metabolite antigens to MAIT cells. Deletion of these chaperones reduces the ERresident MR1 pool and hampers antigen presentation and MAIT cell activation. The MR1 antigen-presentation pathway thus co-opts ER chaperones to fulfill its unique ability to present exogenous metabolite antigens captured within the ER.",

keywords = "Fluorescent probe, MAIT cells, MHC class I-related protein 1 (MR1), Protein trafficking, Vitamin B",

author = "McWilliam, {Hamish E. G.} and Mak, {Jeffrey Y. W.} and Wael Awad and Matthew Zorkau and Sebastian Cruz-Gomez and Lim, {Hui Jing} and Yuting Yan and Sam Wormald and Dagley, {Laura F.} and Eckle, {Sidonia B. G.} and Corbett, {Alexandra J.} and Haiyin Liu and Shihan Li and Reddiex, {Scott J. J.} and Mintern, {Justine D.} and Ligong Liu and James McCluskey and Jamie Rossjohn and Fairlie, {David P.} and Villadangos, {Jose A.}",

year = "2020",

month = oct,

day = "6",

doi = "10.1073/pnas.2011260117",

language = "English",

volume = "117",

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journal = "Proceedings of the National Academy of Sciences of the United States of America",

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McWilliam, HEG, Mak, JYW, Awad, W, Zorkau, M, Cruz-Gomez, S, Lim, HJ, Yan, Y, Wormald, S, Dagley, LF, Eckle, SBG, Corbett, AJ, Liu, H, Li, S, Reddiex, SJJ, Mintern, JD, Liu, L, McCluskey, J, Rossjohn, J, Fairlie, DP & Villadangos, JA 2020, 'Endoplasmic reticulum chaperones stabilize ligand-receptive MR1 molecules for efficient presentation of metabolite antigens', Proceedings of the National Academy of Sciences of the United States of America, vol. 117, no. 40, pp. 24974-24985. https://doi.org/10.1073/pnas.2011260117

Endoplasmic reticulum chaperones stabilize ligand-receptive MR1 molecules for efficient presentation of metabolite antigens. / McWilliam, Hamish E. G.; Mak, Jeffrey Y. W.; Awad, Wael et al.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 117, No. 40, 06.10.2020, p. 24974-24985.

Research output: Contribution to journal › Article › Research › peer-review

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AU - McWilliam, Hamish E. G.

AU - Mak, Jeffrey Y. W.

AU - Awad, Wael

AU - Zorkau, Matthew

AU - Cruz-Gomez, Sebastian

AU - Lim, Hui Jing

AU - Yan, Yuting

AU - Wormald, Sam

AU - Dagley, Laura F.

AU - Eckle, Sidonia B. G.

AU - Corbett, Alexandra J.

AU - Liu, Haiyin

AU - Li, Shihan

AU - Reddiex, Scott J. J.

AU - Mintern, Justine D.

AU - Liu, Ligong

AU - McCluskey, James

AU - Rossjohn, Jamie

AU - Fairlie, David P.

AU - Villadangos, Jose A.

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AB - The antigen-presenting molecule MR1 (MHC class I-related protein 1) presents metabolite antigens derived from microbial vitamin B2 synthesis to activate mucosal-associated invariant T (MAIT) cells. Key aspects of this evolutionarily conserved pathway remain uncharacterized, including where MR1 acquires ligands and what accessory proteins assist ligand binding. We answer these questions by using a fluorophore-labeled stable MR1 antigen analog, a conformation-specific MR1 mAb, proteomic analysis, and a genome-wide CRISPR/Cas9 library screen. We show that the endoplasmic reticulum (ER) contains a pool of two unliganded MR1 conformers stabilized via interactions with chaperones tapasin and tapasin-related protein. This pool is the primary source of MR1 molecules for the presentation of exogenous metabolite antigens to MAIT cells. Deletion of these chaperones reduces the ERresident MR1 pool and hampers antigen presentation and MAIT cell activation. The MR1 antigen-presentation pathway thus co-opts ER chaperones to fulfill its unique ability to present exogenous metabolite antigens captured within the ER.

KW - Fluorescent probe

KW - MAIT cells

KW - MHC class I-related protein 1 (MR1)

KW - Protein trafficking

KW - Vitamin B

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JF - Proceedings of the National Academy of Sciences of the United States of America

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Endoplasmic reticulum chaperones stabilize ligand-receptive MR1 molecules for efficient presentation of metabolite antigens

Abstract

Keywords

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ARC Centre of Excellence in Advanced Molecular Imaging

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Endoplasmic reticulum chaperones stabilize ligand-receptive MR1 molecules for efficient presentation of metabolite antigens

Abstract

Keywords

Access to Document

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Projects

ARC Centre of Excellence in Advanced Molecular Imaging

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