The cytoprotective protein MANF promotes neuronal survival independently from its role as a GRP78 cofactor

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RESEARCH PRODUCT

The cytoprotective protein MANF promotes neuronal survival independently from its role as a GRP78 cofactor

Vera Kovaleva Richard Zimmermann Mart Saarma Maarit Hellman Li-ying Yu Helka Göös Päivi Lindholm Perttu Permi Ave Eesmaa Kristofer Nõges Martin Jung Markku Varjosalo

subject

0301 basic medicine BiFC bimolecular fluorescence complementation MST microscale thermophoresis PDIA1 protein disulfide isomerase family A member 1 Apoptosis NEUROTROPHIC FACTOR MANF Endoplasmic Reticulum Biochemistry protein-protein interaction Mice Bimolecular fluorescence complementation UPR unfolded protein response ENDOPLASMIC-RETICULUM STRESS Mesencephalon Neurotrophic factors Insulin-Secreting Cells Protein Interaction Mapping BINDING COMPREHENSIVE RESOURCE ATF6 unfolded protein response (UPR)PDIA6 protein disulfide isomerase family A member 6 PPIs protein-protein interactions Endoplasmic Reticulum Chaperone BiP Heat-Shock Proteins NPTN neuroplastin biology Chemistry apoptosis unfolded protein response dopamine neurons 3. Good health Cell biology GDNF glial cell line–derived neurotrophic factor IRE1-ALPHA SBD substrate-binding domain endoplasmic reticulum stress MANF mesencephalic astrocyte-derived neurotrophic factor Tm tunicamycin neuroprotection Research Article Protein Binding Signal Transduction GRP78 Protein Disulfide-Isomerase Family Cell Survival TH tyrosine hydroxylase Primary Cell Culture SCG superior cervical ganglion Protein Disulfide-Isomerases IRE1 inositol-requiring enzyme 1 ER-STRESS ER endoplasmic reticulum 03 medical and health sciences ohjelmoitunut solukuolema C-MANF C-terminal domain of MANF CSPs chemical shift perturbations Animals Humans HSP70 Heat-Shock Proteins Nerve Growth Factors NBD nucleotide-binding domain NMR nuclear magnetic resonance Molecular Biology 030102 biochemistry & molecular biology BIP ATF6 Dopaminergic Neurons Gene Expression Profiling Binding protein neuronal cell death DISSOCIATION Cell Biology NEI nucleotide exchange inhibitor Embryo Mammalian adenosiinitrifosfaatti ATP hermosolut mesencephalic astrocyte-derived neurotrophic factor protein–protein interaction PERK protein kinase RNA-like ER kinase HEK293 Cells 030104 developmental biology Gene Expression Regulation Chaperone (protein)Tg thapsigargin biology.protein Unfolded protein response AP-MS affinity purification mass spectrometry 1182 Biochemistry cell and molecular biology GFP-SH SH-tagged GFP endoplasmic reticulum stress (ER stress)DA dopamine mesencephalic astrocyte-derived neurotrophic factor (MANF)proteiinit Neuroplastin

description

Mesencephalic astrocyte-derived neurotrophic factor (MANF) is an endoplasmic reticulum (ER)-stress-regulated protein exhibiting cytoprotective properties through a poorly understood mechanism in various in vitro and in vivo models of neuronal and non-neuronal damage. Although initially characterized as a secreted neurotrophic factor for midbrain dopamine neurons, MANF has recently gained more interest for its intracellular role in regulating the ER homeostasis, including serving as a cofactor of the chaperone glucose-regulated protein 78 (GRP78). We aimed for a better understanding of the neuroprotective mechanisms of MANF. Here we show for the first time that MANF promotes the survival of ER-stressed neurons in vitro as a general unfolded protein response (UPR) regulator, affecting several UPR pathways simultaneously. Interestingly, MANF does not affect naive neurons. We hypothesize that MANF regulates UPR signaling toward a mode more compatible with neuronal survival. Screening of MANF interacting proteins from two mammalian cell lines revealed a conserved interactome of 15 proteins including several ER chaperones such as GRP78, GRP170, protein disulfide isomerase family A member 1, and protein disulfide isomerase family A member 6. Further characterization confirmed previously published finding that MANF is a cofactor of GRP78 interacting with its nucleotide binding domain. Using microscale thermophoresis and nuclear magnetic resonance spectroscopy, we discovered that MANF is an ATP binding protein and that ATP blocks the MANF-GRP78 interaction. Interestingly, functional analysis of the antiapoptotic properties of MANF mutants in cultured neurons revealed divergent roles of MANF as a GRP78 cofactor and as an antiapoptotic regulator of UPR. We conclude that the co-factor type interaction with GRP78 is dispensable for the survival-promoting activity of MANF in neurons. Peer reviewed

year	journal	country	edition	language
2021-01-01

http://urn.fi/URN:NBN:fi:jyu-202102221722