TY - JOUR
T1 - ATM loss disrupts the autophagy-lysosomal pathway
AU - Cheng, Aifang
AU - Tse, Kai Hei
AU - Chow, Hei Man
AU - Gan, Yunqiao
AU - Song, Xuan
AU - Ma, Fulin
AU - Qian, Yi Xuan Yvonne
AU - She, Weiyi
AU - Herrup, Karl
N1 - Funding Information:
We thank Dr. Qian Cai, Dr. Ron Hart and Dr. Mark R. Plummer from Rutgers, the State University of New Jersey, for their helpful advices on lysosome trafficking analysis. We thank APCF, the animal facility in HKUST for animal breeding and Ms. Jianping Ren for animal genotyping. This work was supported by the Research Grants Council, Hong Kong Special Administrative Region (Grants HKUST12/CRF/13G, GRF660813, GRF16101315, AoE/M-05/12, GRF16100718, GRF16103317 and GRF16100219); the Offices of Provost, Vice-President for Research and Graduate Studies, and Dean of Science, The Hong Kong University of Science and Technology (HKUST; Grant VPRGO12SC02); RGC/HKUST Initiation Grant (IGN16SC02), CUHK Improvement on Competitiveness in Hiring New Faculties Start-Up Fund (Ref.:133) and Alzheimer’s Association (AARF-17-531566). We also thank the funding provided by Hong Kong Branch of South Marine Science and Engineering Guangdong Laboratory (SMSEGL20Sc01) and the China Ocean Mineral Resources Research and Development Association (COMRRDA17SC01).
Funding Information:
This work was supported by the Alzheimer?s Association [AARF-17-531566]; Chinese University of Hong Kong [Improvement on Competitiveness in Hiring New Faculties Start-Up Fund - 133]; Research Grants Council, Hong Kong [GRF16101315]; Research Grants Council, Hong Kong SAR [GRF16100219]; Research Grants Council, Hong Kong SAR [IGN16SC02]; Research Grants Council, Hong Kong SAR [GRF16100718]; The Research Grants Council, Hong Kong SAR [GRF660813]; Research Grants Council, Hong Kong SAR [HKUST12/CRF/13G]; Research Grants Council, Hong Kong SAR [AoE/M-05/12]. We thank Dr. Qian Cai, Dr. Ron Hart and Dr. Mark R. Plummer from Rutgers, the State University of New Jersey, for their helpful advices on lysosome trafficking analysis. We thank APCF, the animal facility in HKUST for animal breeding and Ms. Jianping Ren for animal genotyping. This work was supported by the Research Grants Council, Hong Kong Special Administrative Region (Grants HKUST12/CRF/13G, GRF660813, GRF16101315, AoE/M-05/12, GRF16100718, GRF16103317 and GRF16100219); the Offices of Provost, Vice-President for Research and Graduate Studies, and Dean of Science, The Hong Kong University of Science and Technology (HKUST; Grant VPRGO12SC02); RGC/HKUST Initiation Grant (IGN16SC02), CUHK Improvement on Competitiveness in Hiring New Faculties Start-Up Fund (Ref.:133) and Alzheimer?s Association (AARF-17-531566). We also thank the funding provided by Hong Kong Branch of South Marine Science and Engineering Guangdong Laboratory (SMSEGL20Sc01) and the China Ocean Mineral Resources Research and Development Association (COMRRDA17SC01).
Publisher Copyright:
© 2020 Informa UK Limited, trading as Taylor & Francis Group.
PY - 2021/8
Y1 - 2021/8
N2 - ATM (ataxia telangiectasia mutated) protein is found associated with multiple organelles including synaptic vesicles, endosomes and lysosomes, often in cooperation with ATR (ataxia telangiectasia and Rad3 related). Mutation of the ATM gene results in ataxia-telangiectasia (A-T), an autosomal recessive disorder with defects in multiple organs including the nervous system. Precisely how ATM deficiency leads to the complex phenotypes of A-T, however, remains elusive. Here, we reported that part of the connection may lie in autophagy and lysosomal abnormalities. We found that ATM was degraded through the autophagy pathway, while ATR was processed by the proteasome. Autophagy and lysosomal trafficking were both abnormal in atm
−/− neurons and the deficits impacted cellular functions such as synapse maintenance, neuronal survival and glucose uptake. Upregulated autophagic flux was observed in atm
−/− lysosomes, associated with a more acidic pH. Significantly, we found that the ATP6V1A (ATPase, H+ transporting, lysosomal V1 subunit A) proton pump was an ATM kinase target. In atm
−/− neurons, lysosomes showed enhanced retrograde transport and accumulated in the perinuclear regions. We attributed this change to an unexpected physical interaction between ATM and the retrograde transport motor protein, dynein. As a consequence, SLC2A4/GLUT4 (solute carrier family 4 [facilitated glucose transporter], member 4) translocation to the plasma membrane was inhibited and trafficking to the lysosomes was increased, leading to impaired glucose uptake capacity. Together, these data underscored the involvement of ATM in a variety of neuronal vesicular trafficking processes, offering new and therapeutically useful insights into the pathogenesis of A-T. Abbreviations: 3-MA: 3-methyladenine; A-T: ataxia-telangiectasia; ALG2: asparagine-linked glycosylation 2 (alpha-1,3-mannosyltransferase); AMPK: adenosine 5‘-monophosphate (AMP)-activated protein kinase; ATG5: autophagy related 5; ATM: ataxia telangiectasia mutated; ATP6V1A: ATPase, H+ transporting, lysosomal V1 subunit A; ATR: ataxia-telangiectasia and Rad3 related; BFA1: bafilomycin A
1; CC3: cleaved-CASP3; CGN: cerebellar granule neuron; CLQ: chloroquine; CN: neocortical neuron; CTSB: cathepsin B; CTSD: cathepsin D; DYNLL1: the light chain1 of dynein; EIF4EBP1/4E-BP1: eukaryotic translation initiation factor 4E binding protein 1; Etop: etoposide; FBS: fetal bovine serum; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; HBS: HEPES-buffered saline; HEPES: 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; HOMER1: homer protein homolog 1; KU: KU-60019; LAMP1: lysosomal-associated membrane protein 1; LC3B-II: LC3-phosphatidylethanolamine conjugate; Lyso: lysosome; LysopH-GFP: lysopHluorin-GFP; MAP1LC3B/LC3B: microtubule-associated protein 1 light chain 3 beta; MAP2: microtubule associated protein 2; MAPK14: mitogen-activated protein kinase 14; MAPK8/JNK1: mitogen-activated protein kinase 8; MCOLN1/TRPML1: mucolipin 1; OSBPL1A: oxysterol binding protein like 1A; PIKK: phosphatidylinositol 3 kinase related kinase; Rapa: rapamycin; RILP: rab interacting lysosomal protein; ROS: reactive oxygen species; SEM: standard error of mean; SLC2A4/GLUT4: solute carrier family 2 (facilitated glucose transporter), member 4; TSC2/tuberin: TSC complex subunit 2; ULK1: unc-51 like kinase 1; UPS: ubiquitin-proteasome system; VE: VE-822; WCL: whole-cell lysate; WT: wild type.
AB - ATM (ataxia telangiectasia mutated) protein is found associated with multiple organelles including synaptic vesicles, endosomes and lysosomes, often in cooperation with ATR (ataxia telangiectasia and Rad3 related). Mutation of the ATM gene results in ataxia-telangiectasia (A-T), an autosomal recessive disorder with defects in multiple organs including the nervous system. Precisely how ATM deficiency leads to the complex phenotypes of A-T, however, remains elusive. Here, we reported that part of the connection may lie in autophagy and lysosomal abnormalities. We found that ATM was degraded through the autophagy pathway, while ATR was processed by the proteasome. Autophagy and lysosomal trafficking were both abnormal in atm
−/− neurons and the deficits impacted cellular functions such as synapse maintenance, neuronal survival and glucose uptake. Upregulated autophagic flux was observed in atm
−/− lysosomes, associated with a more acidic pH. Significantly, we found that the ATP6V1A (ATPase, H+ transporting, lysosomal V1 subunit A) proton pump was an ATM kinase target. In atm
−/− neurons, lysosomes showed enhanced retrograde transport and accumulated in the perinuclear regions. We attributed this change to an unexpected physical interaction between ATM and the retrograde transport motor protein, dynein. As a consequence, SLC2A4/GLUT4 (solute carrier family 4 [facilitated glucose transporter], member 4) translocation to the plasma membrane was inhibited and trafficking to the lysosomes was increased, leading to impaired glucose uptake capacity. Together, these data underscored the involvement of ATM in a variety of neuronal vesicular trafficking processes, offering new and therapeutically useful insights into the pathogenesis of A-T. Abbreviations: 3-MA: 3-methyladenine; A-T: ataxia-telangiectasia; ALG2: asparagine-linked glycosylation 2 (alpha-1,3-mannosyltransferase); AMPK: adenosine 5‘-monophosphate (AMP)-activated protein kinase; ATG5: autophagy related 5; ATM: ataxia telangiectasia mutated; ATP6V1A: ATPase, H+ transporting, lysosomal V1 subunit A; ATR: ataxia-telangiectasia and Rad3 related; BFA1: bafilomycin A
1; CC3: cleaved-CASP3; CGN: cerebellar granule neuron; CLQ: chloroquine; CN: neocortical neuron; CTSB: cathepsin B; CTSD: cathepsin D; DYNLL1: the light chain1 of dynein; EIF4EBP1/4E-BP1: eukaryotic translation initiation factor 4E binding protein 1; Etop: etoposide; FBS: fetal bovine serum; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; HBS: HEPES-buffered saline; HEPES: 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; HOMER1: homer protein homolog 1; KU: KU-60019; LAMP1: lysosomal-associated membrane protein 1; LC3B-II: LC3-phosphatidylethanolamine conjugate; Lyso: lysosome; LysopH-GFP: lysopHluorin-GFP; MAP1LC3B/LC3B: microtubule-associated protein 1 light chain 3 beta; MAP2: microtubule associated protein 2; MAPK14: mitogen-activated protein kinase 14; MAPK8/JNK1: mitogen-activated protein kinase 8; MCOLN1/TRPML1: mucolipin 1; OSBPL1A: oxysterol binding protein like 1A; PIKK: phosphatidylinositol 3 kinase related kinase; Rapa: rapamycin; RILP: rab interacting lysosomal protein; ROS: reactive oxygen species; SEM: standard error of mean; SLC2A4/GLUT4: solute carrier family 2 (facilitated glucose transporter), member 4; TSC2/tuberin: TSC complex subunit 2; ULK1: unc-51 like kinase 1; UPS: ubiquitin-proteasome system; VE: VE-822; WCL: whole-cell lysate; WT: wild type.
KW - Ataxia-telangiectasia
KW - autophagy
KW - lysosome
KW - neurodegeneration
KW - protein degradation
KW - SLC2A4/GLUT4
KW - trafficking
UR - http://www.scopus.com/inward/record.url?scp=85089546989&partnerID=8YFLogxK
U2 - 10.1080/15548627.2020.1805860
DO - 10.1080/15548627.2020.1805860
M3 - Journal article
C2 - 32757690
AN - SCOPUS:85089546989
VL - 17
SP - 1998
EP - 2010
JO - Autophagy
JF - Autophagy
SN - 1554-8627
IS - 8
ER -