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Change of DVL3 interacting partners upon its polyglutamylation

ABSTRACT: Polyglutamylation is a reversible post-translational modification initially discovered on tubulin. In eukaryotes, polyglutamylation is catalyzed by enzymes from the tubulin tyrosine ligase-like (TTLL) family, which show substrate and reaction specificities that are so far mostly explored on tubulin. By contrast, little is known on the mechanism and significance of TTLL-mediated modification of non-tubulin substrates. Here, we found that TTLL11 generates a previously unknown type of polyglutamylation by adding glutamate residues to the carboxy-terminus of a substrate protein. TTLL11 efficiently polyglutamylates the signaling protein disheveled 3 (DVL3), thereby changing the interactome of DVL3, as well as its capacity to undergo liquid-liquid phase separation (LLPS). Both carboxyterminal polyglutamylation and the resulting reduction in LLPS capacity of DVL3 were reverted by the deglutamylating enzyme CCP6, which demonstrates the causal relationship between TTLL11-mediated polyglutamylation and LLPS. We thus discovered a novel type of posttranslational modification catalyzed by a TTLL enzyme, which significantly broadens the range of proteins that can be modified by polyglutamylation, and provide first evidence that polyglutamylation can act as a regulator of protein LLPS.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: David Potesil

LAB HEAD: Zbynek Zdrahal

PROVIDER: PXD033548 | Pride | 2024-09-30

REPOSITORIES: Pride

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			Action	DRS
	DVL3-WT-TTLL11_01.raw	Raw
	DVL3-WT-TTLL11_02.raw	Raw
	DVL3-WT-TTLL11_03.raw	Raw
	DVL3-WT_01.raw	Raw
	DVL3-WT_02.raw	Raw

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Analysis of DVL3 polyglutamylation using Q-TOF

Project description:Polyglutamylation is a reversible post-translational modification initially discovered on tubulin. In eukaryotes, polyglutamylation is catalyzed by enzymes from the tubulin tyrosine ligase-like (TTLL) family, which show substrate and reaction specificities that are so far mostly explored on tubulin. By contrast, little is known on the mechanism and significance of TTLL-mediated modification of non-tubulin substrates. Here, we found that TTLL11 generates a previously unknown type of polyglutamylation by adding glutamate residues to the carboxy-terminus of a substrate protein. TTLL11 efficiently polyglutamylates the signaling protein disheveled 3 (DVL3), thereby changing the interactome of DVL3, as well as its capacity to undergo liquid-liquid phase separation (LLPS). Both carboxyterminal polyglutamylation and the resulting reduction in LLPS capacity of DVL3 were reverted by the deglutamylating enzyme CCP6, which demonstrates the causal relationship between TTLL11-mediated polyglutamylation and LLPS. We thus discovered a novel type of posttranslational modification catalyzed by a TTLL enzyme, which significantly broadens the range of proteins that can be modified by polyglutamylation, and provide first evidence that polyglutamylation can act as a regulator of protein LLPS.

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Disruption of Ttll5/Stamp gene (Tubulin tyrosine ligase-like protein 5/SRC-1 and TIF2 associated modulatory protein gene) in male mice causes sperm malformation and infertility

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