Cargando…

Structural and functional studies of the first tripartite protein complex at the Trypanosoma brucei flagellar pocket collar

The flagellar pocket (FP) is the only endo- and exocytic organelle in most trypanosomes and, as such, is essential throughout the life cycle of the parasite. The neck of the FP is maintained enclosed around the flagellum via the flagellar pocket collar (FPC). The FPC is a macromolecular cytoskeletal...

Descripción completa

Detalles Bibliográficos
Autores principales: Isch, Charlotte, Majneri, Paul, Landrein, Nicolas, Pivovarova, Yulia, Lesigang, Johannes, Lauruol, Florian, Robinson, Derrick R., Dong, Gang, Bonhivers, Mélanie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8360560/
https://www.ncbi.nlm.nih.gov/pubmed/34339455
http://dx.doi.org/10.1371/journal.ppat.1009329
_version_ 1783737768465137664
author Isch, Charlotte
Majneri, Paul
Landrein, Nicolas
Pivovarova, Yulia
Lesigang, Johannes
Lauruol, Florian
Robinson, Derrick R.
Dong, Gang
Bonhivers, Mélanie
author_facet Isch, Charlotte
Majneri, Paul
Landrein, Nicolas
Pivovarova, Yulia
Lesigang, Johannes
Lauruol, Florian
Robinson, Derrick R.
Dong, Gang
Bonhivers, Mélanie
author_sort Isch, Charlotte
collection PubMed
description The flagellar pocket (FP) is the only endo- and exocytic organelle in most trypanosomes and, as such, is essential throughout the life cycle of the parasite. The neck of the FP is maintained enclosed around the flagellum via the flagellar pocket collar (FPC). The FPC is a macromolecular cytoskeletal structure and is essential for the formation of the FP and cytokinesis. FPC biogenesis and structure are poorly understood, mainly due to the lack of information on FPC composition. To date, only two FPC proteins, BILBO1 and FPC4, have been characterized. BILBO1 forms a molecular skeleton upon which other FPC proteins can, theoretically, dock onto. We previously identified FPC4 as the first BILBO1 interacting partner and demonstrated that its C-terminal domain interacts with the BILBO1 N-terminal domain (NTD). Here, we report by yeast two-hybrid, bioinformatics, functional and structural studies the characterization of a new FPC component and BILBO1 partner protein, BILBO2 (Tb927.6.3240). Further, we demonstrate that BILBO1 and BILBO2 share a homologous NTD and that both domains interact with FPC4. We have determined a 1.9 Å resolution crystal structure of the BILBO2 NTD in complex with the FPC4 BILBO1-binding domain. Together with mutational analyses, our studies reveal key residues for the function of the BILBO2 NTD and its interaction with FPC4 and evidenced a tripartite interaction between BILBO1, BILBO2, and FPC4. Our work sheds light on the first atomic structure of an FPC protein complex and represents a significant step in deciphering the FPC function in Trypanosoma brucei and other pathogenic kinetoplastids.
format Online
Article
Text
id pubmed-8360560
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-83605602021-08-13 Structural and functional studies of the first tripartite protein complex at the Trypanosoma brucei flagellar pocket collar Isch, Charlotte Majneri, Paul Landrein, Nicolas Pivovarova, Yulia Lesigang, Johannes Lauruol, Florian Robinson, Derrick R. Dong, Gang Bonhivers, Mélanie PLoS Pathog Research Article The flagellar pocket (FP) is the only endo- and exocytic organelle in most trypanosomes and, as such, is essential throughout the life cycle of the parasite. The neck of the FP is maintained enclosed around the flagellum via the flagellar pocket collar (FPC). The FPC is a macromolecular cytoskeletal structure and is essential for the formation of the FP and cytokinesis. FPC biogenesis and structure are poorly understood, mainly due to the lack of information on FPC composition. To date, only two FPC proteins, BILBO1 and FPC4, have been characterized. BILBO1 forms a molecular skeleton upon which other FPC proteins can, theoretically, dock onto. We previously identified FPC4 as the first BILBO1 interacting partner and demonstrated that its C-terminal domain interacts with the BILBO1 N-terminal domain (NTD). Here, we report by yeast two-hybrid, bioinformatics, functional and structural studies the characterization of a new FPC component and BILBO1 partner protein, BILBO2 (Tb927.6.3240). Further, we demonstrate that BILBO1 and BILBO2 share a homologous NTD and that both domains interact with FPC4. We have determined a 1.9 Å resolution crystal structure of the BILBO2 NTD in complex with the FPC4 BILBO1-binding domain. Together with mutational analyses, our studies reveal key residues for the function of the BILBO2 NTD and its interaction with FPC4 and evidenced a tripartite interaction between BILBO1, BILBO2, and FPC4. Our work sheds light on the first atomic structure of an FPC protein complex and represents a significant step in deciphering the FPC function in Trypanosoma brucei and other pathogenic kinetoplastids. Public Library of Science 2021-08-02 /pmc/articles/PMC8360560/ /pubmed/34339455 http://dx.doi.org/10.1371/journal.ppat.1009329 Text en © 2021 Isch et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Isch, Charlotte
Majneri, Paul
Landrein, Nicolas
Pivovarova, Yulia
Lesigang, Johannes
Lauruol, Florian
Robinson, Derrick R.
Dong, Gang
Bonhivers, Mélanie
Structural and functional studies of the first tripartite protein complex at the Trypanosoma brucei flagellar pocket collar
title Structural and functional studies of the first tripartite protein complex at the Trypanosoma brucei flagellar pocket collar
title_full Structural and functional studies of the first tripartite protein complex at the Trypanosoma brucei flagellar pocket collar
title_fullStr Structural and functional studies of the first tripartite protein complex at the Trypanosoma brucei flagellar pocket collar
title_full_unstemmed Structural and functional studies of the first tripartite protein complex at the Trypanosoma brucei flagellar pocket collar
title_short Structural and functional studies of the first tripartite protein complex at the Trypanosoma brucei flagellar pocket collar
title_sort structural and functional studies of the first tripartite protein complex at the trypanosoma brucei flagellar pocket collar
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8360560/
https://www.ncbi.nlm.nih.gov/pubmed/34339455
http://dx.doi.org/10.1371/journal.ppat.1009329
work_keys_str_mv AT ischcharlotte structuralandfunctionalstudiesofthefirsttripartiteproteincomplexatthetrypanosomabruceiflagellarpocketcollar
AT majneripaul structuralandfunctionalstudiesofthefirsttripartiteproteincomplexatthetrypanosomabruceiflagellarpocketcollar
AT landreinnicolas structuralandfunctionalstudiesofthefirsttripartiteproteincomplexatthetrypanosomabruceiflagellarpocketcollar
AT pivovarovayulia structuralandfunctionalstudiesofthefirsttripartiteproteincomplexatthetrypanosomabruceiflagellarpocketcollar
AT lesigangjohannes structuralandfunctionalstudiesofthefirsttripartiteproteincomplexatthetrypanosomabruceiflagellarpocketcollar
AT lauruolflorian structuralandfunctionalstudiesofthefirsttripartiteproteincomplexatthetrypanosomabruceiflagellarpocketcollar
AT robinsonderrickr structuralandfunctionalstudiesofthefirsttripartiteproteincomplexatthetrypanosomabruceiflagellarpocketcollar
AT donggang structuralandfunctionalstudiesofthefirsttripartiteproteincomplexatthetrypanosomabruceiflagellarpocketcollar
AT bonhiversmelanie structuralandfunctionalstudiesofthefirsttripartiteproteincomplexatthetrypanosomabruceiflagellarpocketcollar