Identification of TbPBN1 in Trypanosoma brucei reveals a conserved heterodimeric architecture for glycosylphosphatidylinositol-mannosyltransferase-I.
Trypanosoma brucei
Endoplasmic reticulum (ER)
glycosylphosphatidylinositol (GPI)
glycosyltransferase
mannosyltransferase
Journal
Molecular microbiology
ISSN: 1365-2958
Titre abrégé: Mol Microbiol
Pays: England
ID NLM: 8712028
Informations de publication
Date de publication:
02 2022
02 2022
Historique:
revised:
06
12
2021
received:
01
11
2021
accepted:
06
12
2021
pubmed:
8
12
2021
medline:
5
4
2022
entrez:
7
12
2021
Statut:
ppublish
Résumé
Glycosylphosphatidylinositol (GPI)-anchored proteins are found in all eukaryotes and are especially abundant on the surface of protozoan parasites such as Trypanosoma brucei. GPI-mannosyltransferase-I (GPI-MT-I) catalyzes the addition of the first of three mannoses that make up the glycan core of GPI. Mammalian and yeast GPI-MT-I consist of two essential subunits, the catalytic subunit PIG-M/Gpi14 and the accessory subunit PIG-X/Pbn1(mammals/yeast). T. brucei GPI-MT-I has been highlighted as a potential antitrypanosome drug target but has not been fully characterized. Here, we show that T. brucei GPI-MT-I also has two subunits, TbGPI14 and TbPBN1. Using TbGPI14 deletion, and TbPBN1 RNAi-mediated depletion, we show that both proteins are essential for the mannosyltransferase activity needed for GPI synthesis and surface expression of GPI-anchored proteins. In addition, using native PAGE and co-immunoprecipitation analyses, we demonstrate that TbGPI14 and TbPBN1 interact to form a higher-order complex. Finally, we show that yeast Gpi14 does not restore GPI-MT-I function in TbGPI14 knockout trypanosomes, consistent with previously demonstrated species specificity within GPI-MT-I subunit associations. The identification of an essential trypanosome GPI-MT-I subcomponent indicates wide conservation of the heterodimeric architecture unusual for a glycosyltransferase, leaving open the question of the role of the noncatalytic TbPBN1 subunit in GPI-MT-I function.
Identifiants
pubmed: 34875117
doi: 10.1111/mmi.14859
pmc: PMC9306709
doi:
Substances chimiques
Glycosylphosphatidylinositols
0
Protozoan Proteins
0
Mannosyltransferases
EC 2.4.1.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
450-461Informations de copyright
© 2021 The Authors. Molecular Microbiology published by John Wiley & Sons Ltd.
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