Structural basis of tubulin tyrosination by tubulin tyrosine ligase

Prota, A.E.; Magiera, Maria M.; Kuijpers, Marijn; Bargsten, Katja; Frey, Daniel; Wieser, Mara M.; Jaussi, Rolf; Hoogenraad, Casper C.; Kammerer, Richard A.; Janke, Carsten; Steinmetz, Michel O.

doi:10.1083/jcb.201211017

Cited by 197 publications

(237 citation statements)

References 76 publications

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“…R389 is also invariant. Its equivalent residue in TTL coordinates the tubulin-tail glutamate that ligates to the incoming tyrosine (39). Consistent with their importance in substrate binding, R389E and R411E mutations in TTLL3 reduce glycylation to background levels (Fig.…”

Section: Ttll3 Preferentially Initiates Glycyl Chains On β-Tubulin Tamentioning

confidence: 72%

“…The active site is lined with basic residues conserved in all TTL and TTLL enzymes. Invariant R411 stabilizes the ATP γ-phosphate in TTL (39,40) (Fig. 4 A and C).…”

Section: Ttll3 Preferentially Initiates Glycyl Chains On β-Tubulin Tamentioning

confidence: 99%

“…The structure of the glutamylase TTLL7 (38) revealed how sequence elements specific to glutamylases differentiate the conserved TTL scaffold (39,40). To elucidate what architectural features specialize the TTL fold for glycylation, we determined the X-ray crystal structure of the conserved core of Xenopus tropicalis TTLL3 (residues 6-569) bound to the slowly hydrolyzable ATP analog AMPPNP (Fig.…”

Section: Ttll3 Preferentially Initiates Glycyl Chains On β-Tubulin Tamentioning

confidence: 99%

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Crystal structure of tubulin tyrosine ligase-like 3 reveals essential architectural elements unique to tubulin monoglycylases

Garnham

et al. 2017

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

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Glycylation and glutamylation, the posttranslational addition of glycines and glutamates to genetically encoded glutamates in the intrinsically disordered tubulin C-terminal tails, are crucial for the biogenesis and stability of cilia and flagella and play important roles in metazoan development. Members of the diverse family of tubulin tyrosine ligase-like (TTLL) enzymes catalyze these modifications, which are part of an evolutionarily conserved and complex tubulin code that regulates microtubule interactions with cellular effectors. The site specificity of TTLL enzymes and their biochemical interplay remain largely unknown. Here, we report an in vitro characterization of a tubulin glycylase. We show that TTLL3 glycylates the β-tubulin tail at four sites in a hierarchical order and that TTLL3 and the glutamylase TTLL7 compete for overlapping sites on the tubulin tail, providing a molecular basis for the anticorrelation between glutamylation and glycylation observed in axonemes. This anticorrelation demonstrates how a combinatorial tubulin code written in two different posttranslational modifications can arise through the activities of related but distinct TTLL enzymes. To elucidate what structural elements differentiate TTLL glycylases from glutamylases, with which they share the common TTL scaffold, we determined the TTLL3 X-ray structure at 2.3-Å resolution. This structure reveals two architectural elements unique to glycyl initiases and critical for their activity. Thus, our work sheds light on the structural and functional diversification of TTLL enzymes, and constitutes an initial important step toward understanding how the tubulin code is written through the intersection of activities of multiple TTLL enzymes.tubulin code | TTLL enzymes | glycylation | tubulin modification | glutamylation

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Section: Ttll3 Preferentially Initiates Glycyl Chains On β-Tubulin Tamentioning

confidence: 72%

“…The active site is lined with basic residues conserved in all TTL and TTLL enzymes. Invariant R411 stabilizes the ATP γ-phosphate in TTL (39,40) (Fig. 4 A and C).…”

Section: Ttll3 Preferentially Initiates Glycyl Chains On β-Tubulin Tamentioning

confidence: 99%

Section: Ttll3 Preferentially Initiates Glycyl Chains On β-Tubulin Tamentioning

confidence: 99%

See 1 more Smart Citation

Crystal structure of tubulin tyrosine ligase-like 3 reveals essential architectural elements unique to tubulin monoglycylases

Garnham

et al. 2017

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

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“…These results show both that stronger centromeres are more likely to detach and that the cortical side is more susceptible to detachment. To test whether the enrichment of Tyr α-tubulin makes the cortical side more unstable, we manipulated tyrosination levels by modulating the expression of Tubulin Tyrosine Ligase (TTL), which catalyze α-tubulin tyrosination (33). Increasing Tyr α-tubulin by overexpressing TTL destabilized spindle MTs ( Fig.…”

mentioning

confidence: 99%

Spindle asymmetry drives non-Mendelian chromosome segregation

Akera

Chmátal

Trimm

et al. 2017

Preprint

Self Cite

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Genetic elements compete for transmission through meiosis, when haploid gametes are created from a diploid parent. Selfish elements can enhance their transmission through meiotic drive, in violation of Mendel's Law of Segregation. In female meiosis, selfish elements drive by preferentially attaching to the egg side of the spindle, which implies some asymmetry between the two sides of the spindle, but molecular mechanisms underlying spindle asymmetry are unknown. Here we show that CDC42 signaling from the cell cortex regulates microtubule tyrosination to induce spindle asymmetry, and non-Mendelian segregation depends on this asymmetry. These signals depend on cortical polarization directed by chromosomes, which are positioned near the cortex to allow the asymmetric cell division. Thus, selfish meiotic drivers exploit the asymmetry inherent in female meiosis to bias their transmission.Genetic conflict is inherent in any haploid-diploid life cycle because genetic elements compete for transmission to the offspring through meiosis, the process by which haploids are generated. Mendel's Law of Segregation states that alleles of a gene are transmitted with equal probability, but it is increasingly clear that this law is often violated, and segregation can be manipulated by selfish genetic elements through meiotic drive. Drive can occur by eliminating competing gametes that do not contain the selfish element (e.g., sperm killing or spore killing) or by exploiting the asymmetry in female meiosis to increase the transmission of the selfish element to the egg. Although the impact of meiotic drive on many aspects of evolution and genetics is now recognized, with examples widespread across eukaryotes (1-4), the underlying mechanisms are largely unknown.Female meiosis provides a clear opportunity for selfish elements to cheat because of its inherent asymmetry: only chromosomes that segregate to the egg can be transmitted to offspring, while the rest are degraded in polar bodies. Conceptually, female meiotic drive depends on three conditions: asymmetry in cell fate, a functional not peer-reviewed) is the author/funder. All rights reserved. No reuse allowed without permission.The copyright holder for this preprint (which was . http://dx.doi.org/10.1101/180869 doi: bioRxiv preprint first posted online Aug. 25, 2017; 2 difference between homologous chromosomes that influences their segregation, and asymmetry within the meiotic spindle (5). The asymmetry in cell fate is well established (6), and chromosomal rearrangements and amplifications of repetitive sequences (e.g., centromeres) are associated with biased segregation (7-10). Asymmetry within the meiotic spindle was noted in grasshopper in 1976 (11), but not studied further, and molecular mechanisms regulating such asymmetry are unknown.Oocyte spindles are positioned close to the cortex and oriented perpendicular to the cortex in order to achieve the highly asymmetric cell division, so that cytokinesis produces a large egg and a small polar body (Fig. 1A). A selfish element ...

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“…12 As the TTL uses only soluble tubulin dimers as substrates, detyrosinated microtubules must first be disassembled for re-tyrosination of their dimers before their re-assembly. 13,14 At the molecular level, tubulin tyrosination-detyrosination is a well-known characteristic of microtubule dynamics. 15,16 Stable/longlasting microtubules with a low dynamicity often harbor detyrosinated tubulin, whereas tyrosinated tubulin is mostly found in dynamic microtubules.…”

Section: Introductionmentioning

confidence: 99%

CAP-Gly proteins contribute to microtubule-dependent trafficking via interactions with the C-terminal aromatic residue of α-tubulin

2017

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SUMMARYIn mammals, the C-terminal tyrosine residue of a-tubulin is subjected to removal/re-addition cycles resulting in tyrosinated microtubules and detyrosinated Glu-microtubules. CLIP170 and its yeast ortholog (Bik1) interact weakly with Glu-microtubules. Recently, we described a Microtubule-Rho1-and Bik1-dependent mechanism involved in Snc1 routing. Here, we further show a contribution of the yeast p150Glued ortholog (Nip100) in Snc1 trafficking. Both CLIP170 and p150Glued are CAPGly-containing proteins that belong to the microtubule Cend-tracking protein family (known as CTips). We discuss the CTips-dependent role of microtubules in trafficking, the role of CAP-Gly proteins as possible molecular links between microtubules and vesicles, as well as the contribution of the Rho1-GTPase to the regulation of the CTips repertoire and the partners associated with microtubules.

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Structural basis of tubulin tyrosination by tubulin tyrosine ligase

Abstract: Structural analysis of a complex of tubulin and tubulin tyrosine ligase (TTL) reveals insights into TTL’s enzymatic mechanism, how it discriminates between α- and β-tubulin, and its possible evolutionary origin.

Cited by 197 publications

References 76 publications

Crystal structure of tubulin tyrosine ligase-like 3 reveals essential architectural elements unique to tubulin monoglycylases

Crystal structure of tubulin tyrosine ligase-like 3 reveals essential architectural elements unique to tubulin monoglycylases

Spindle asymmetry drives non-Mendelian chromosome segregation

CAP-Gly proteins contribute to microtubule-dependent trafficking via interactions with the C-terminal aromatic residue of α-tubulin

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