Subpellicular and flagellar microtubules of Trypanosoma brucei brucei contain the same alpha-tubulin isoforms.

Schneider, André; Sherwin, Trevor; Sasse, Rosemary; Russell, David G.; Gull, Keith; Seebeck, Thomas

doi:10.1083/jcb.104.3.431

Cited by 139 publications

(129 citation statements)

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“…This result and the lack of any detyrosinated tx peptide argue that Giardia tubulin is not subject to a detyrosination/tyrosination cycle. Here we note that tx-tubulin of Trypanosoma brucei, a member of the Kinetoplastida, is, in line with its sequence, subject to such a cycle [31,32] while tx-tubulins from some ciliates show sequences not compatible with the normal cycle (see for instance [14]). The reason for this interesting variability is not known.…”

Section: Discussionmentioning

confidence: 92%

Polyglycylation of tubulin in the diplomonad Giardia lamblia, one of the oldest eukaryotes

et al. 1996

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We have searched for post-translational modifications in tubulin of the diplomonad Giardia lamblia, which is a representative of the earliest branches in eukaryotic evolution. The carboxyterminal peptide of a-tubulin was isolated and characterized by automated sequencing and mass spectrometry. Some 60% of the peptide is unmodified, while the remainder shows various degrees of polyglycylation. The number of glycyl residues in the lateral side chain ranges from 2 to 23. All peptide species encountered end with alanine-tyrosine, indicating the absence of a detyrosination/tyrosination cycle. We conclude that tubulin-specific polyglycylation could be as old as tubulin and axonemal structures.Key words: Flagella; Polyglutamylation; Polyglycylation; Post-translational modification; Tubulin; Tyrosination to search for their evolutionary origin because they are unique to tubulin, a typical eukaryotic protein.Based on ultrastructural characteristics and several molecular phylogenies there is general agreement that the diplomonads were among the first branches which emerged from the eukaryotic tree. Diplomonads, like other Archezoa, are thought to have arisen before the acquisition of mitochondria and to have retained many primitive features of the first nucleated cells [17][18][19][20][21]. Giardia lamblia is a particularly wellcharacterized diplomonad. Its cytoskeleton is dominated by microtubules. Giardia has eight flagellar axonemes and a large disc cytoskeleton which consists of rnicrotubules and the microribbons, which arise from ~-giardin [22,23]. Here we report an analysis of the post-translational modifications of ct-tubulin from Giardia based on the characterization of its carboxyterminal peptide.

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Section: Discussionmentioning

confidence: 92%

Polyglycylation of tubulin in the diplomonad Giardia lamblia, one of the oldest eukaryotes

et al. 1996

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“…IF of procyclic 29-13 cell lines was essentially done as described in ref. 42. In short, cells were fixed in PBS containing 4% (wt/vol) paraformaldehyde for 10 min and subsequently permeabilized for 5 min with PBS containing 2% (wt/vol) Triton X-100.…”

Section: Methodsmentioning

confidence: 99%

Trypanosomal TAC40 constitutes a novel subclass of mitochondrial β-barrel proteins specialized in mitochondrial genome inheritance

Schnarwiler

Niemann

Doiron

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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Mitochondria cannot form de novo but require mechanisms allowing their inheritance to daughter cells. In contrast to most other eukaryotes Trypanosoma brucei has a single mitochondrion whose single-unit genome is physically connected to the flagellum. Here we identify a β-barrel mitochondrial outer membrane protein, termed tripartite attachment complex 40 (TAC40), that localizes to this connection. TAC40 is essential for mitochondrial DNA inheritance and belongs to the mitochondrial porin protein family. However, it is not specifically related to any of the three subclasses of mitochondrial porins represented by the metabolite transporter voltage-dependent anion channel (VDAC), the protein translocator of the outer membrane 40 (TOM40), or the fungi-specific MDM10, a component of the endoplasmic reticulum-mitochondria encounter structure (ERMES). MDM10 and TAC40 mediate cellular architecture and participate in transmembrane complexes that are essential for mitochondrial DNA inheritance. In yeast MDM10, in the context of the ERMES, is postulated to connect the mitochondrial genomes to actin filaments, whereas in trypanosomes TAC40 mediates the linkage of the mitochondrial DNA to the basal body of the flagellum. However, TAC40 does not colocalize with trypanosomal orthologs of ERMES components and, unlike MDM10, it regulates neither mitochondrial morphology nor the assembly of the protein translocase. TAC40 therefore defines a novel subclass of mitochondrial porins that is distinct from VDAC, TOM40, and MDM10. However, whereas the architecture of the TAC40-containing complex in trypanosomes and the MDM10-containing ERMES in yeast is very different, both are organized around a β-barrel protein of the mitochondrial porin family that mediates a DNA-cytoskeleton linkage that is essential for mitochondrial DNA inheritance.organelle | parasitic protozoa

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“…Isolation of T. brucei flagella was performed as previously described (18,47). Approximately 2 × 10 9 procyclic 29.13 cells (48) were extracted sequentially with buffers containing 1% Nonidet P-40 and 1 M KCl.…”

Section: Methodsmentioning

confidence: 99%

Structure of Trypanosoma brucei flagellum accounts for its bihelical motion

Koyfman

Schmid

Gheiratmand

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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Trypanosoma brucei is a parasitic protozoan that causes African sleeping sickness. It contains a flagellum required for locomotion and viability. In addition to a microtubular axoneme, the flagellum contains a crystalline paraflagellar rod (PFR) and connecting proteins. We show here, by cryoelectron tomography, the structure of the flagellum in three bending states. The PFR lattice in straight flagella repeats every 56 nm along the length of the axoneme, matching the spacing of the connecting proteins. During flagellar bending, the PFR crystallographic unit cell lengths remain constant while the interaxial angles vary, similar to a jackscrew. The axoneme drives the expansion and compression of the PFR lattice. We propose that the PFR modifies the in-plane axoneme motion to produce the characteristic trypanosome bihelical motility as captured by high-speed light microscope videography.T rypanosoma brucei has devastated the African continent for centuries by infecting humans and domestic animals and has hindered economic development in sub-Saharan Africa (1). Current sleeping sickness treatments are inadequate and the drugs used are highly toxic (2). In recent years, the motility of the T. brucei flagellum has been found to be essential for parasite survival, infection, and disease pathogenesis (3), and has emerged as a promising drug target (4). Flagella with similar structural organization and protein composition have also been found in euglenoids (5) and other kinetoplastid parasites including Leishmania spp. and Trypanosoma cruzi, which cause Leishmaniasis and Chagas disease, respectively (6).The trypanosome flagellum is more complex than most other eukaryotic microtubule-based flagella (7-9) and is completely different from rotary-motor based bacterial flagella (10). Each T. brucei cell contains one flagellum that moves the cell body in an alternating right and left-handed twist resulting in bihelical motion (11) (Movie S1). The membrane-enclosed flagellum, composed of an axoneme, a paraflagellar rod (PFR) (12), and connecting proteins, is attached to the cell body (Fig. 1). PFR was identified as a lattice-like ultrastructure in T. brucei flagellum (13). This periodic and crystalline nature of the PFR was confirmed in T. brucei (14) and related species (15, 16). Monoclonal antibody screens (17) and proteomics studies (18-20) have identified at least 40 PFR proteins. Among them, PFR1 (73 kDa) and PFR2 (69 kDa), containing coiled-coil regions (21), are major structural components of the PFR (22). Depletion of these proteins results in failure of PFR assembly and cell motility defects (17, 23) ( Fig. S1 and Movie S2). In the T. brucei pathogenic bloodstream form, ablation of PFR2 causes death of the parasite (18). These results demonstrate a critical role of the PFR in T. brucei motility and viability. We have employed cryoelectron tomography (cryo-ET) to determine the structure of a biochemically isolated T. brucei flagella (18). We describe here a model that explains how the structure and arrangement of the fl...

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Subpellicular and flagellar microtubules of Trypanosoma brucei brucei contain the same alpha-tubulin isoforms.

Cited by 139 publications

References 50 publications

Polyglycylation of tubulin in the diplomonad Giardia lamblia, one of the oldest eukaryotes

Polyglycylation of tubulin in the diplomonad Giardia lamblia, one of the oldest eukaryotes

Trypanosomal TAC40 constitutes a novel subclass of mitochondrial β-barrel proteins specialized in mitochondrial genome inheritance

Structure of Trypanosoma brucei flagellum accounts for its bihelical motion

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