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...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2021-08-01
|
| Series: | PLoS Pathogens |
| Online Access: | https://doi.org/10.1371/journal.ppat.1009329 |
| _version_ | 1818574153716858880 |
|---|---|
| author | Charlotte Isch Paul Majneri Nicolas Landrein Yulia Pivovarova Johannes Lesigang Florian Lauruol Derrick R Robinson Gang Dong Mélanie Bonhivers |
| author_facet | Charlotte Isch Paul Majneri Nicolas Landrein Yulia Pivovarova Johannes Lesigang Florian Lauruol Derrick R Robinson Gang Dong Mélanie Bonhivers |
| author_sort | Charlotte Isch |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-15T00:22:41Z |
| format | Article |
| id | doaj.art-342552844d3f4eaabbb926a00273446c |
| institution | Directory Open Access Journal |
| issn | 1553-7366 1553-7374 |
| language | English |
| last_indexed | 2024-12-15T00:22:41Z |
| publishDate | 2021-08-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Pathogens |
| spelling | doaj.art-342552844d3f4eaabbb926a00273446c2022-12-21T22:42:15ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742021-08-01178e100932910.1371/journal.ppat.1009329Structural and functional studies of the first tripartite protein complex at the Trypanosoma brucei flagellar pocket collar.Charlotte IschPaul MajneriNicolas LandreinYulia PivovarovaJohannes LesigangFlorian LauruolDerrick R RobinsonGang DongMélanie BonhiversThe 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.https://doi.org/10.1371/journal.ppat.1009329 |
| spellingShingle | Charlotte Isch Paul Majneri Nicolas Landrein Yulia Pivovarova Johannes Lesigang Florian Lauruol Derrick R Robinson Gang Dong Mélanie Bonhivers Structural and functional studies of the first tripartite protein complex at the Trypanosoma brucei flagellar pocket collar. PLoS Pathogens |
| 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 |
| url | https://doi.org/10.1371/journal.ppat.1009329 |
| work_keys_str_mv | AT charlotteisch structuralandfunctionalstudiesofthefirsttripartiteproteincomplexatthetrypanosomabruceiflagellarpocketcollar AT paulmajneri structuralandfunctionalstudiesofthefirsttripartiteproteincomplexatthetrypanosomabruceiflagellarpocketcollar AT nicolaslandrein structuralandfunctionalstudiesofthefirsttripartiteproteincomplexatthetrypanosomabruceiflagellarpocketcollar AT yuliapivovarova structuralandfunctionalstudiesofthefirsttripartiteproteincomplexatthetrypanosomabruceiflagellarpocketcollar AT johanneslesigang structuralandfunctionalstudiesofthefirsttripartiteproteincomplexatthetrypanosomabruceiflagellarpocketcollar AT florianlauruol structuralandfunctionalstudiesofthefirsttripartiteproteincomplexatthetrypanosomabruceiflagellarpocketcollar AT derrickrrobinson structuralandfunctionalstudiesofthefirsttripartiteproteincomplexatthetrypanosomabruceiflagellarpocketcollar AT gangdong structuralandfunctionalstudiesofthefirsttripartiteproteincomplexatthetrypanosomabruceiflagellarpocketcollar AT melaniebonhivers structuralandfunctionalstudiesofthefirsttripartiteproteincomplexatthetrypanosomabruceiflagellarpocketcollar |