Tubulin polyglutamylase: isozymic variants and regulation during the cell cycle in HeLa cells.
Polyglutamylation is a posttranslational modification of tubulin that is very common in neurons and ciliated or flagellated cells. It was proposed to regulate the binding of microtubule associated proteins (MAPs) and molecular motors as a function of the length of the polyglutamyl side-chain. Though much less common, this modification of tubulin also occurs in proliferating cells like HeLa cells where it is associated with centrioles and with the mitotic spindle. Recently, we partially purified tubulin polyglutamylase from mouse brain and described its enzymatic properties. In this work, we focused on tubulin polyglutamylase activity from HeLa cells. Our results support the existence of a tubulin polyglutamylase family composed of several isozymic variants specific for alpha- or beta-tubulin subunits. In the latter case, the specificity probably also concerns the different beta-tubulin isotypes. Interestingly, we found that tubulin polyglutamylase activity is regulated in a cell cycle dependent manner and peaks in G(2)-phase while the level of glutamylated tubulin peaks in mitosis. Consistent results were obtained by treating the cells with hydroxyurea, nocodazole or taxotere. In particular, in mitotic cells, tubulin polyglutamylase activity was always low while glutamylation level was high. Finally, tubulin polyglutamylase activity and the level of glutamylated tubulin appeared to be inversely related. This paradox suggests a complex regulation of both tubulin polyglutamylase and the reverse deglutamylase activity.
Regnard
C
C
Laboratoire de Biochimie Cellulaire, CNRS UPR 9065 and Université Paris VI, Collège de France, 75005 Paris. regnard@ext.jussieu.fr
Desbruyères
E
E
Denoulet
P
P
Eddé
B
B
D000818:Animals; D001921:Brain / Q000201:enzymology; D002453:Cell Cycle; D002455:Cell Division; D002502:Centrioles / Q000201:enzymology; D000077143:Docetaxel; D016195:G2 Phase; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D014644:Genetic Variation; D006367:HeLa Cells; D006801:Humans; D006918:Hydroxyurea / Q000494:pharmacology; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D051379:Mice; D015739:Nocodazole / Q000494:pharmacology; D017239:Paclitaxel / Q000031:analogs & derivatives / Q000494:pharmacology; D010453:Peptide Synthases; D011099:Polyglutamic Acid / Q000235:genetics* / Q000378:metabolism; D008941:Spindle Apparatus / Q000201:enzymology; D043823:Taxoids; D014404:Tubulin / Q000235:genetics* / Q000378:metabolism
D007527:Isoenzymes; D043823:Taxoids; D014404:Tubulin; D000077143:Docetaxel; D011099:Polyglutamic Acid; D010453:Peptide Synthases; C112698:tubulin polyglutamylase; D017239:Paclitaxel; D015739:Nocodazole; D006918:Hydroxyurea