Polyglycylation of Tubulin: a Posttranslational Modification in Axonemal Microtubules

Redeker, Virginie; Levilliers, Nicolette; Schmitter, Jean Marie; Caer, JP Le; Rossier, Jean; Adoutte, André; Bré, Marie‐Hélène

doi:10.1126/science.7992051

Cited by 260 publications

(247 citation statements)

References 31 publications

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“…The exact position of this modified residue has been identified in all brain tubulins [3,8,[10][11][12][13]. The more recently discovered polyglycylation has been documented for ct-and 13-tubulin from the ciliary axonemal microtubules of the protist Paramecium [14] and from the sperm axonemal microtubules of mammals [15] (Plessmann, U. and Weber, K. unpublished results) and sea urchins [16]. Thus all post-translational modifications unique to tubulin involve the negatively charged carboxyterminal 10 to 15 residues [12,15].…”

Section: Introduction 2 Materials and Methodsmentioning

confidence: 96%

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: Introduction 2 Materials and Methodsmentioning

confidence: 96%

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

et al. 1996

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“…Many PTMs involve the reversible addition of a range of molecules of variable size that range from methyl, phosphate, and acetyl groups, to lipids and small proteins. Glutamylation [1] and glycylation [2] are unusual PTMs, as the amino acids are added as homopolymeric chains of differing length that are covalently attached to a side-chain carboxyl group of individual glutamic acid residues near the carboxyl terminus of specific proteins. These modifications were originally discovered in tubulins.…”

Section: Introductionmentioning

confidence: 99%

TTLL10 is a protein polyglycylase that can modify nucleosome assembly protein 1

et al. 2008

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Certain proteins can undergo polyglycylation and polyglutamylation. Polyglutamylases (glutamate ligases) have recently been identified in a family of tubulin tyrosine ligase-like (TTLL) proteins. However, no polyglycylase (glycine ligase) has yet been reported. Here we identify a polyglycylase in the TTLL proteins by using an anti-poly-glycine antibody. The antibody reacted with a cytoplasmic 60-kDa protein that accumulated in elongating spermatids. Using tandem mass spectrometry of trypsinized samples, immunoprecipitated by the antibody from the TTLL10-expressing cells, we identified the 60-kDa protein as nucleosome assembly protein 1 (NAP1). Recombinant TTLL10 incorporated glycine into recombinant NAP1 in vitro. Mutational analyses indicated that Glu residues at 359 and 360 in the C-terminal part of NAP1 are putative sites for the modification. Thus, TTLL10 is a polyglycylase for NAP1.

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“…Tubulin glycylation is conserved in protists and most metazoans (17). The glycyl chain length on either α-or β-tubulin tails in axonemes is highly variable (16)(17)(18), ranging from 1 to 40 posttranslationally added glycines (19). Of the 13 TTLL enzymes in mammals (10, 11), three are glycylases: TTLL3, -8, and -10 (12,13,20).…”

mentioning

confidence: 99%

“…Glycylation is the ATP-dependent addition of glycines, either singly or in chains, to internal glutamates (16). Tubulin glycylation is conserved in protists and most metazoans (17).…”

mentioning

confidence: 99%

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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Polyglycylation of Tubulin: a Posttranslational Modification in Axonemal Microtubules

Cited by 260 publications

References 31 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

TTLL10 is a protein polyglycylase that can modify nucleosome assembly protein 1

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

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