Polyglutamylation is a post-translational modification with a broad range of substrates.
Polyglutamylation is a post-translational modification that generates lateral acidic side chains on proteins by sequential addition of glutamate amino acids. This modification was first discovered on tubulins, and it is important for several microtubule functions. Besides tubulins, only the nucleosome assembly proteins NAP1 and NAP2 have been shown to be polyglutamylated. Here, using a proteomic approach, we identify a large number of putative substrates for polyglutamylation in HeLa cells. By analyzing a selection of these putative substrates, we show that several of them can serve as in vitro substrates for two of the recently discovered polyglutamylases, TTLL4 and TTLL5. We further show that TTLL4 is the main polyglutamylase enzyme present in HeLa cells and that new substrates of polyglutamylation are indeed modified by TTLL4 in a cellular context. No clear consensus polyglutamylation site could be defined from the primary sequence of the here-identified new substrates of polyglutamylation. However, we demonstrate that glutamate-rich stretches are important for a protein to become polyglutamylated. Most of the newly identified substrates of polyglutamylation are nucleocytoplasmic shuttling proteins, including many chromatin-binding proteins. Our work reveals that polyglutamylation is a much more widespread post-translational modification than initially thought and thus that it might be a regulator of many cellular processes.
van Dijk
Juliette
J
Centre de Recherche de Biochimie Macromoléculaire, Universités Montpellier 2 and 1, CNRS, 34293 Montpellier, France.
Miro
Julie
J
Strub
Jean-Marc
JM
Lacroix
Benjamin
B
van Dorsselaer
Alain
A
Edde
Bernard
B
Janke
Carsten
C
D000681:Amylases / Q000378:metabolism; D015153:Blotting, Western; D002846:Chromatography, Affinity; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D011099:Polyglutamic Acid / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D013379:Substrate Specificity
D011099:Polyglutamic Acid; D000681:Amylases