Polyglutamylase activity of tubulin tyrosine ligase-like 4 is negatively regulated by the never in mitosis gene A family kinase never in mitosis gene A-related kinase 5  

作　　者：Talita Diniz Melo-Hanchuk Jörg Kobarg 

机构地区：[1]Faculty of Pharmaceutical Sciences,Unicamp,Campinas 13083-862,Brazil [2]Faculty of Pharmaceutical Sciences,University of Campinas,Campinas 13083-862,Brazil

出　　处：《World Journal of Biological Chemistry》2021年第3期38-51,共14页世界生物化学杂志（英文版）（电子版）

基　　金：Fundação de AmparoàPesquisa do Estado São Paulo(FAPESP,São Paulo,Brazil)through Grant Temático,No.2017/03489-1.

摘　　要：BACKGROUND Tubulins,building blocks of microtubules,are modified substrates of diverse post-translational modifications including phosphorylation,polyglycylation and polyglutamylation.Polyglutamylation of microtubules,catalyzed by enzymes from the tubulin tyrosine ligase-like(TTLL)family,can regulate interactions with molecular motors and other proteins.Due to the diversity and functional importance of microtubule modifications,strict control of the TTLL enzymes has been suggested.AIM To characterize the interaction between never in mitosis gene A-related kinase 5(NEK5)and TTLL4 proteins and the effects of TTLL4 phosphorylation.METHODS The interaction between NEK5 and TTLL4 was identified by yeast two-hybrid screening using the C-terminus of NEK5(a.a.260–708)as bait and confirmed by immunoprecipitation.The phosphorylation sites of TTLL4 were identified by mass spectrometry and point mutations were introduced.RESULTS Here,we show that NEK5 interacts with TTLL4 and regulates its polyglutamylation activity.We further show that NEK5 can also interact with TTLL5 and TTLL7.The silencing of NEK5 increases the levels of polyglutamylation of proteins by increasing the activity of TTLL4.The same effects were observed after the expression of the catalytically inactive form of NEK5.This regulation of TTLL4 activity involves its phosphorylation at Y815 and S1136 amino acid residues.CONCLUSION Our results demonstrate,for the first time,the regulation of TTLL activity through phosphorylation,pointing to NEK5 as a potential effector kinase.We also suggest a general control of tubulin polyglutamylation through NEK family members in human cells.

关 键 词：KINASE Polyglutamylation Never in mitosis gene A-related kinase 5 Tubulin tyrosine ligase-like 4 Microtubules Post translational regulation 

分 类 号：Q253[生物学—细胞生物学]