Protein Kinase A Rescues Microtubule Affinity-regulating Kinase 2-induced Microtubule Instability and Neurite Disruption by Phosphorylating Serine 409

Deng, Si‐Ping; Wu, Lichen; Wang, Ya-Chao; Cao, Peng-Rong; Xu, Lei; Li, Qianru; Li, Meng; Zhang, Lun; Jiang, Yuejing; Yang, Xiaoyu; Sun, Shengnan; Tan, Minjia; Qian, Min; Zang, Yi; Feng, Linyin; Li, Jia

doi:10.1074/jbc.m114.629873

Cited by 9 publications

(6 citation statements)

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“…Overexpression of MARK 2/4 inhibited the activities of both CRTC2 and to a lesser extent CRTC3 ( Fig 6A ). Despite the presence of a consensus PKA phosphorylation site at S409 ( Fig 6B ), which has been reported to modulate MARK2 activity [ 35 ], phospho-mimetic and phospho-incompetent mutations at S409 had no effect on MARK2-mediated phosphorylation of CRTC2/3 at sites II and III ( Fig 6C ). We confirmed that S409 is indeed phosphorylated in response to Fsk stimulation by using a phospho-PKA substrate antibody ( Fig 6D ).…”

Section: Resultsmentioning

confidence: 91%

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Analysis of a cAMP regulated coactivator family reveals an alternative phosphorylation motif for AMPK family members

et al. 2017

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The second messenger cAMP stimulates cellular gene expression via the PKA-mediated phosphorylation of the transcription factor CREB and through dephosphorylation of the cAMP-responsive transcriptional coactivators (CRTCs). Under basal conditions, CRTCs are phosphorylated by members of the AMPK family of Ser/Thr kinases and sequestered in the cytoplasm via a phosphorylation-dependent association with 14-3-3 proteins. Increases in cAMP promote the dephosphorylation and nuclear translocation of CRTCs, where they bind to CREB and stimulate relevant target genes. Although they share considerable sequence homology, members of the CRTC family exert non-overlapping effects on cellular gene expression through as yet unidentified mechanisms. Here we show that the three CRTCs exhibit distinct patterns of 14-3-3 binding at three conserved sites corresponding to S70, S171, and S275 (in CRTC2). S171 functions as the gatekeeper site for 14-3-3 binding; it acts cooperatively with S275 in stabilizing this interaction following its phosphorylation by the cAMP-responsive SIK and the cAMP-nonresponsive MARK kinases. Although S171 contains a consensus recognition site for phosphorylation by AMPK family members, S70 and S275 carry variant motifs (MNTGGS275LPDL), lacking basic residues that are otherwise critical for SIK/MARK recognition as well as 14-3-3 binding. Correspondingly, the activity of these motifs differs between CRTC family members. As the variant (SLPDL) motif is present and apparently phosphorylated in other mammalian proteins, our studies suggest that the regulation of cellular targets by AMPK family members is more extensive than previously appreciated.

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Section: Resultsmentioning

confidence: 91%

“…cAMP has been reported to inhibit SIKs and other AMPK family members through PKA-mediated phosphorylation [ 1 , 35 ]. Indeed, exposure to Fsk decreased endogenous CRTC2 and CRTC3 phosphorylation at sites II and III ( Fig 5A ).…”

Section: Resultsmentioning

confidence: 99%

Analysis of a cAMP regulated coactivator family reveals an alternative phosphorylation motif for AMPK family members

et al. 2017

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“…It has been demonstrated that active MARK2 interacted with tau and phosphorylated tau at Ser262 in stably transfected cell line, while the interaction between MARK2 and tau was significantly reduced when MARK2 was inhibited by protein kinase inhibitor [25]. The overexpression of MARK2 led to hyperphosphorylation of MAPs and disturbance in microtubule array, resulting in cellular morphological changes and cell death in Chinese hamster ovary (CHO) cells [1926], while knockdown of MARK2 expression using siRNA contributed to the formation of multiple axon like neurites and elongation of axons in hippocampal neurons [2627]. One study reported that MARK2 and PTEN-induced kinase 1 (PINK1) were co-localized with mitochondrial and regulate their transport, and MARK2 was identified as an upstream regulator of PINK1 [28].…”

Section: Discussionmentioning

confidence: 99%

“…Previous studies have reported that NMDAR activated protein kinase A (PKA) which phosphorylated MARK2 [29]. MARK2 could regulate microtubule stability, neurite outgrowth, and early phosphorylation of tau in AD [26]. The augmented interactions between MARK2 and tau in AD brain suggest that MARK2 plays an important role in the early phosphorylation of tau in AD, indicating it as a potential therapeutic target.…”

Section: Discussionmentioning

confidence: 99%

Memantine Improves Cognitive Function and Alters Hippocampal and Cortical Proteome in Triple Transgenic Mouse Model of Alzheimer's Disease

et al. 2019

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Memantine is a non-competitive N-methyl-D-aspartate receptor (NMDAR) antagonist clinically approved for moderate-to-severe Alzheimer' s disease (AD) to improve cognitive functions. There is no report about the proteomic alterations induced by memantine in AD mouse model yet. In this study, we investigated the protein profiles in the hippocampus and the cerebral cortex of AD-related transgenic mouse model (3×Tg-AD) treated with memantine. Mice (8-month) were treated with memantine (5 mg/kg/bid) for 4 months followed by behavioral and molecular evaluation. Using step-down passive avoidance (SDA) test, novel object recognition (NOR) test and Morris water maze (MWM) test, it was observed that memantine significantly improved learning and memory retention in 3xTg-AD mice. By using quantitative proteomic analysis, 3301 and 3140 proteins in the hippocampus and the cerebral cortex respectively were identified to be associated with AD abnormalities. In the hippocampus, memantine significantly altered the expression levels of 233 proteins, among which PCNT, ATAXIN2, TNIK, and NOL3 were up-regulated, and FLNA, MARK 2 and BRAF were down-regulated. In the cerebral cortex, memantine significantly altered the expression levels of 342 proteins, among which PCNT, PMPCB, CRK, and MBP were up-regulated, and DNM2, BRAF, TAGLN 2 and FRY1 were down-regulated. Further analysis with bioinformatics showed that memantine modulated biological pathways associated with cytoskeleton and ErbB signaling in the hippocampus, and modulated biological pathways associated with axon guidance, ribosome, cytoskeleton, calcium and MAPK signaling in the cerebral cortex. Our data indicate that memantine induces higher levels of proteomic alterations in the cerebral cortex than in the hippocampus, suggesting memantine affects various brain regions in different manners. Our study provides a novel view on the complexity of protein responses induced by memantine in the brain of AD.

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“…There are four members of the Par1/MARK family in mammals. Par1/MARK plays an instrumental role in regulating various cellular processes in the brain including neuronal migration [22–24], neurite outgrowth [25], dendritic spine formation, and plasticity [26–31]. However, whether and how Par1 affects microglia function has yet to be explored.…”

Section: Introductionmentioning

confidence: 99%

Loss of Par1b/MARK2 primes microglia during brain development and enhances their sensitivity to injury

DiBona

Zhu

Shah

et al. 2019

J Neuroinflammation

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BackgroundMicroglia, the resident immune cells of the brain, exhibit various morphologies that correlate with their functions under physiological and pathological conditions. In conditions such as aging and stress, microglia priming occurs, which leads to altered morphology and lower threshold for activation upon further insult. However, the molecular mechanisms that lead to microglia priming are unclear.MethodsTo understand the role of Par1b/MARK2 in microglia, we first expressed shRNA targeting luciferase or Par1b/MARK2 in primary microglial cells and imaged the cells using fluorescent microscopy to analyze for morphological changes. A phagocytosis assay was then used to assess functional changes. We then moved in vivo and used a Par1b/MARK2 knockout mouse model to assess for changes in microglia density, morphology, and phagocytosis using immunohistochemistry, confocal imaging, and 3D image reconstruction. Next, we used two-photon in vivo imaging in live Par1b/MARK2 deficient mice to examine microglia dynamics. In addition, a controlled-cortical impact injury was performed on wild-type and Par1b/MARK2-deficient mice and microglial response was determined by confocal imaging. Finally, to help rule out non-cell autonomous effects, we analyzed apoptosis by confocal imaging, cytokine levels by multiplex ELISA, and blood-brain barrier permeability using Evans Blue assay.ResultsHere, we show that loss of the cell polarity protein Par1b/MARK2 facilitates the activation of primary microglia in culture. We next found that microglia in Par1b/MARK2 deficient mice show increased density and a hypertrophic morphology. These morphological changes are accompanied with alterations in microglia functional responses including increased phagocytosis of neuronal particles early in development and decreased surveillance of the brain parenchyma, all reminiscent of a primed phenotype. Consistent with this, we found that microglia in Par1b/MARK2 deficient mice have a significantly lower threshold for activation upon injury.ConclusionsTogether, our studies show that loss of Par1b/MARK2 switches microglia from a surveillant to a primed state during development, resulting in an increased neuroinflammatory response to insults.Electronic supplementary materialThe online version of this article (10.1186/s12974-018-1390-3) contains supplementary material, which is available to authorized users.

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Protein Kinase A Rescues Microtubule Affinity-regulating Kinase 2-induced Microtubule Instability and Neurite Disruption by Phosphorylating Serine 409

Cited by 9 publications

References 48 publications

Analysis of a cAMP regulated coactivator family reveals an alternative phosphorylation motif for AMPK family members

Analysis of a cAMP regulated coactivator family reveals an alternative phosphorylation motif for AMPK family members

Memantine Improves Cognitive Function and Alters Hippocampal and Cortical Proteome in Triple Transgenic Mouse Model of Alzheimer's Disease

Loss of Par1b/MARK2 primes microglia during brain development and enhances their sensitivity to injury

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