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Ubiquitin-like protein 3 (UBL3) is required for MARCH ubiquitination of major histocompatibility complex class II and CD86

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posted on 2022-05-10, 00:00 authored by H Liu, KR Wilson, AM Firth, C Macri, P Schriek, AB Blum, J Villar, S Wormald, Mitch ShambrookMitch Shambrook, B Xu, HJ Lim, HEG McWilliam, Andrew HillAndrew Hill, LE Edgington-Mitchell, I Caminschi, MH Lahoud, E Segura, MJ Herold, JA Villadangos, JD Mintern
The MARCH E3 ubiquitin (Ub) ligase MARCH1 regulates trafficking of major histocompatibility complex class II (MHC II) and CD86, molecules of critical importance to immunity. Here we show, using a genome-wide CRISPR knockout screen, that ubiquitin-like protein 3 (UBL3) is a necessary component of ubiquitination-mediated trafficking of these molecules in mice and in humans. Ubl3-deficient mice have elevated MHC II and CD86 expression on the surface of professional and atypical antigen presenting cells. UBL3 also regulates MHC II and CD86 in human dendritic cells (DCs) and macrophages. UBL3 impacts ubiquitination of MARCH1 substrates, a mechanism that requires UBL3 plasma membrane anchoring via prenylation. Loss of UBL3 alters adaptive immunity with impaired development of thymic regulatory T cells, loss of conventional type 1 DCs, increased number of trogocytic marginal zone B cells, and defective in vivo MHC II and MHC I antigen presentation. In summary, we identify UBL3 as a conserved, critical factor in MARCH1-mediated ubiquitination with important roles in immune responses.


We thank the Antibody Services Facility and Genomics Hub (Walter and Eliza Hall Institute) for the provision of reagents and expert assistance. We acknowledge the Melbourne Cytometry Platform from the Bio21 Institute node (University of Melbourne) for the provision of flow cytometry services. We acknowledge the Melbourne Mass Spectrometry and Proteomics Facility of the Bio21 Molecular Science and Biotechnology Institute at the University of Melbourne for the support of mass spectrometry analysis. We thank Satoshi Ishido (Hyogo College of Medicine, Nishinomiya, Japan) for providing the Marchf1-/- and Marchf8-/- mice. The generation of Ubl3-/- mice by CRISPR/Cas9 was performed by Andrew Kueh and Marco Herold (WEHI MAGEC laboratory). This work was supported by National Health and Medical Research Council, Australia grants and fellowships (1058193, 1154502, 1113293, and 1163090 [to J.A.V.], 1161101 and 1129672 [to J.D.M.], 2003192 [to H.E.G.McW], GNT1132604 [to A.F.H.], 1159658, 1186575, and 1156095 [to M.J.H.], and 1142358 [to M.H.L.]), Australian Research Council, Department of Education and Training grants or fellowships (110101383, 190102213, 170102471, 160103134 [to J.A.V.] and 190101242, 180100844, 160101373, and 180100521 [to J.D.M.], DE180100418 [to L.E.-M.]), the Leukemia and Lymphoma Society of America (LLS SCOR 7015-18 to M.J.H.), the Cancer Council of Victoria (1147328 and 2021 Grant In Aid to M.J.H.), and a Human Frontiers Science Program grant (0064/2011 [to J.A.V.]).


Publication Date



Nature Communications





Article Number

ARTN 1934




Springer Nature



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