The transmembrane domain and luminal C-terminal region independently support invariant chain trimerization and assembly with MHCII into nonamers.
Antigen Presentation
Antigens, Differentiation, B-Lymphocyte
/ genetics
Cell Membrane
/ metabolism
HEK293 Cells
HLA-A24 Antigen
/ metabolism
HLA-DR Antigens
/ genetics
Histocompatibility Antigens Class II
/ genetics
Humans
Mutagenesis, Site-Directed
Mutation
/ genetics
Protein Domains
/ genetics
Protein Multimerization
Antigen presentation
CD74
MHCII
Nonamerization
RXR
Transmembrane domain
Trimerization domain
Journal
BMC immunology
ISSN: 1471-2172
Titre abrégé: BMC Immunol
Pays: England
ID NLM: 100966980
Informations de publication
Date de publication:
12 08 2021
12 08 2021
Historique:
received:
25
08
2020
accepted:
20
07
2021
entrez:
13
8
2021
pubmed:
14
8
2021
medline:
29
1
2022
Statut:
epublish
Résumé
Invariant chain (CD74, Ii) is a multifunctional protein expressed in antigen presenting cells. It assists the ER exit of various cargos and serves as a receptor for the macrophage migration inhibitory factor. The newly translated Ii chains trimerize, a structural feature that is not readily understood in the context of its MHCII chaperoning function. Two segments of Ii, the luminal C-terminal region (TRIM) and the transmembrane domain (TM), have been shown to participate in the trimerization process but their relative importance and impact on the assembly with MHCII molecules remains debated. Here, we addressed the requirement of these domains in the trimerization of human Ii as well as in the oligomerization with MHCII molecules. We used site-directed mutagenesis to generate series of Ii and DR mutants. These were transiently transfected in HEK293T cells to test their cell surface expression and analyse their interactions by co-immunoprecipitations. Our results showed that the TRIM domain is not essential for Ii trimerization nor for intracellular trafficking with MHCII molecules. We also gathered evidence that in the absence of TM, TRIM allows the formation of multi-subunit complexes with HLA-DR. Similarly, in the absence of TRIM, Ii can assemble into high-order structures with MHCII molecules. Altogether, our data show that trimerization of Ii through either TM or TRIM sustains nonameric complex formation with MHCII molecules.
Sections du résumé
BACKGROUND
Invariant chain (CD74, Ii) is a multifunctional protein expressed in antigen presenting cells. It assists the ER exit of various cargos and serves as a receptor for the macrophage migration inhibitory factor. The newly translated Ii chains trimerize, a structural feature that is not readily understood in the context of its MHCII chaperoning function. Two segments of Ii, the luminal C-terminal region (TRIM) and the transmembrane domain (TM), have been shown to participate in the trimerization process but their relative importance and impact on the assembly with MHCII molecules remains debated. Here, we addressed the requirement of these domains in the trimerization of human Ii as well as in the oligomerization with MHCII molecules. We used site-directed mutagenesis to generate series of Ii and DR mutants. These were transiently transfected in HEK293T cells to test their cell surface expression and analyse their interactions by co-immunoprecipitations.
RESULTS
Our results showed that the TRIM domain is not essential for Ii trimerization nor for intracellular trafficking with MHCII molecules. We also gathered evidence that in the absence of TM, TRIM allows the formation of multi-subunit complexes with HLA-DR. Similarly, in the absence of TRIM, Ii can assemble into high-order structures with MHCII molecules.
CONCLUSIONS
Altogether, our data show that trimerization of Ii through either TM or TRIM sustains nonameric complex formation with MHCII molecules.
Identifiants
pubmed: 34384367
doi: 10.1186/s12865-021-00444-6
pii: 10.1186/s12865-021-00444-6
pmc: PMC8362237
doi:
Substances chimiques
Antigens, Differentiation, B-Lymphocyte
0
HLA-A*24:02 antigen
0
HLA-A24 Antigen
0
HLA-DR Antigens
0
Histocompatibility Antigens Class II
0
invariant chain
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
56Informations de copyright
© 2021. The Author(s).
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