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PMID:20519502

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Citation

Kimura, Y, Kurabe, N, Ikegami, K, Tsutsumi, K, Konishi, Y, Kaplan, OI, Kunitomo, H, Iino, Y, Blacque, OE and Setou, M (2010) Identification of tubulin deglutamylase among Caenorhabditis elegans and mammalian cytosolic carboxypeptidases (CCPs). J. Biol. Chem. 285:22936-41

Abstract

Tubulin polyglutamylation is a reversible post-translational modification, serving important roles in microtubule (MT)-related processes. Polyglutamylases of the tubulin tyrosine ligase-like (TTLL) family add glutamate moieties to specific tubulin glutamate residues, whereas as yet unknown deglutamylases shorten polyglutamate chains. First we investigated regulatory machinery of tubulin glutamylation in MT-based sensory cilia of the roundworm Caenorhabditis elegans. We found that ciliary MTs were polyglutamylated by a process requiring ttll-4. Conversely, loss of ccpp-6 gene function, which encodes one of two cytosolic carboxypeptidases (CCPs), resulted in elevated levels of ciliary MT polyglutamylation. Consistent with a deglutamylase function for ccpp-6, overexpression of this gene in ciliated cells decreased polyglutamylation signals. Similarly, we confirmed that overexpression of murine CCP5, one of two sequence orthologs of nematode ccpp-6, caused a dramatic loss of MT polyglutamylation in cultured mammalian cells. Finally, using an in vitro assay for tubulin glutamylation, we found that recombinantly expressed Myc-tagged CCP5 exhibited deglutamylase biochemical activities. Together, these data from two evolutionarily divergent systems identify C. elegans CCPP-6 and its mammalian ortholog CCP5 as a tubulin deglutamylase.

Links

PubMed PMC2906286 Online version:10.1074/jbc.C110.128280

Keywords

Animals; Caenorhabditis elegans/cytology; Caenorhabditis elegans/enzymology; Caenorhabditis elegans/genetics; Caenorhabditis elegans/metabolism; Carboxypeptidases/chemistry; Carboxypeptidases/genetics; Carboxypeptidases/metabolism; Cell Line; Cilia/metabolism; Cytosol/enzymology; Humans; Mice; Microtubules/metabolism; Peptide Synthases/chemistry; Peptide Synthases/genetics; Peptide Synthases/metabolism; Protein Processing, Post-Translational; Sequence Homology, Amino Acid

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

CAEEL:CBPC6

involved_in

GO:0035608: protein deglutamylation

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

CAEEL:TTLL4

involved_in

GO:0018095: protein polyglutamylation

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

MOUSE:CBPC5

enables

GO:0004181: metallocarboxypeptidase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

MOUSE:CBPC5

involved_in

GO:0035611: protein branching point deglutamylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

MOUSE:CBPC5

involved_in

GO:0035608: protein deglutamylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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