TAU is a microtubule-associated protein that promotes microtubule assembly and stability in the axon.TAU is missorted and aggregated in an array of diseases known as tauopathies.Microtubules are essential for neuronal...TAU is a microtubule-associated protein that promotes microtubule assembly and stability in the axon.TAU is missorted and aggregated in an array of diseases known as tauopathies.Microtubules are essential for neuronal function and regulated via a complex set of post-translational modifications,changes of which affect microtubule stability and dynamics,microtubule interaction with other proteins and cellular structures,and mediate recruitment of microtubule-severing enzymes.As impairment of microtubule dynamics causes neuronal dysfunction,we hypothesize cognitive impairment in human disease to be impacted by impairment of microtubule dynamics.We therefore aimed to study the effects of a disease-causing mutation of TAU(P301L)on the levels and localization of microtubule post-translational modifications indicative of microtubule stability and dynamics,to assess whether P301L-TAU causes stability-changing modifications to microtubules.To investigate TAU localization,phosphorylation,and effects on tubulin post-translational modifications,we expressed wild-type or P301L-TAU in human MAPT-KO induced pluripotent stem cell-derived neurons(i Neurons)and studied TAU in neurons in the hippocampus of mice transgenic for human P301L-TAU(p R5 mice).Human neurons expressing the longest TAU isoform(2N4R)with the P301L mutation showed increased TAU phosphorylation at the AT8,but not the p-Ser-262 epitope,and increased polyglutamylation and acetylation of microtubules compared with endogenous TAU-expressing neurons.P301L-TAU showed pronounced somatodendritic presence,but also successful axonal enrichment and a similar axodendritic distribution comparable to exogenously expressed 2N4R-wildtype-TAU.P301L-TAU-expressing hippocampal neurons in transgenic mice showed prominent missorting and tauopathy-typical AT8-phosphorylation of TAU and increased polyglutamylation,but reduced acetylation,of microtubules compared with non-transgenic littermates.In sum,P301L-TAU results in changes in microtubule PTMs,suggestive of impairment of microtubule stability.This is accompanied by missorting and aggregation of TAU in mice but not in i Neurons.Microtubule PTMs/impairment may be of key importance in tauopathies.展开更多
类PR5受体蛋白激酶(PR5 receptor-like protein kinase,PR5Ks)包含病程相关蛋白5(PR5)和激酶结构域,为植物类受体蛋白激酶(Receptor-like protein kinase,RLK)家族的1个亚族,与植物的抗病防御反应相关。本研究从小麦基因组中鉴定了17个P...类PR5受体蛋白激酶(PR5 receptor-like protein kinase,PR5Ks)包含病程相关蛋白5(PR5)和激酶结构域,为植物类受体蛋白激酶(Receptor-like protein kinase,RLK)家族的1个亚族,与植物的抗病防御反应相关。本研究从小麦基因组中鉴定了17个PR5Ks基因,对其理化性质、基因结构、保守基序和启动子进行分析。结果表明,17个PR5Ks蛋白家族成员均为疏水性蛋白,氨基酸数目在415~663个,分子量在44.60~73.52 kD,等电点介于5.61~8.4,蛋白结构以无规则卷曲和α螺旋为主。顺式作用元件分析显示,在小麦PR5Ks基因转录起始位点上游2000 bp序列中存在多种响应元件,包括SA、MeJA、厌氧诱导、低温等响应元件。为了明确17个小麦PR5Ks成员是否具有抗病性,对小麦在赤霉菌(Fusarium graminearum)和条锈菌(Puccinia striiformis f.sp.tritici)侵染过程中PR5Ks基因的表达规律进行分析,发现TaPR5K-3、TaPR5K-5、TaPR5K-9和TaPR5K-11受赤霉菌和条锈菌的共同诱导。利用实时荧光定量PCR(Quantitative real-time PCR,qPCR)技术检测了接种叶锈菌的小麦中基因的表达模式,结果显示,TaPR5K-3、TaPR5K-5、TaPR5K-9和TaPR5K-11这4个基因均受叶锈菌诱导且在抗病植株中高表达,表明他们在小麦与叶锈菌的互作中发挥正向调控作用。综上所述,本研究初步筛选到4个与小麦抗病相关的PR5Ks基因,为进一步揭示PR5Ks蛋白家族参与小麦生长发育及抗病防御反应提供参考。展开更多
基金supported by the Koeln Fortune Program/Faculty of Medicine,University of Cologne,the Alzheimer Forschung Initiative e.V.(grant#22039,to HZ)open-access funding from the DFG/GRC issued to the University of CologneAlzheimer Forschung Initiative e.V.for Open Access Publishing(a publication grant#P2401,to MAAK)。
文摘TAU is a microtubule-associated protein that promotes microtubule assembly and stability in the axon.TAU is missorted and aggregated in an array of diseases known as tauopathies.Microtubules are essential for neuronal function and regulated via a complex set of post-translational modifications,changes of which affect microtubule stability and dynamics,microtubule interaction with other proteins and cellular structures,and mediate recruitment of microtubule-severing enzymes.As impairment of microtubule dynamics causes neuronal dysfunction,we hypothesize cognitive impairment in human disease to be impacted by impairment of microtubule dynamics.We therefore aimed to study the effects of a disease-causing mutation of TAU(P301L)on the levels and localization of microtubule post-translational modifications indicative of microtubule stability and dynamics,to assess whether P301L-TAU causes stability-changing modifications to microtubules.To investigate TAU localization,phosphorylation,and effects on tubulin post-translational modifications,we expressed wild-type or P301L-TAU in human MAPT-KO induced pluripotent stem cell-derived neurons(i Neurons)and studied TAU in neurons in the hippocampus of mice transgenic for human P301L-TAU(p R5 mice).Human neurons expressing the longest TAU isoform(2N4R)with the P301L mutation showed increased TAU phosphorylation at the AT8,but not the p-Ser-262 epitope,and increased polyglutamylation and acetylation of microtubules compared with endogenous TAU-expressing neurons.P301L-TAU showed pronounced somatodendritic presence,but also successful axonal enrichment and a similar axodendritic distribution comparable to exogenously expressed 2N4R-wildtype-TAU.P301L-TAU-expressing hippocampal neurons in transgenic mice showed prominent missorting and tauopathy-typical AT8-phosphorylation of TAU and increased polyglutamylation,but reduced acetylation,of microtubules compared with non-transgenic littermates.In sum,P301L-TAU results in changes in microtubule PTMs,suggestive of impairment of microtubule stability.This is accompanied by missorting and aggregation of TAU in mice but not in i Neurons.Microtubule PTMs/impairment may be of key importance in tauopathies.
