Alzheimer Disease-specific Conformation of Hyperphosphorylated Paired Helical Filament-Tau Is Polyubiquitinated through Lys-48, Lys-11, and Lys-6 Ubiquitin Conjugation

Department of Pharmaceutical Sciences, University of Southern California, Los Angeles, California 90089, USA.
Journal of Biological Chemistry (Impact Factor: 4.57). 05/2006; 281(16):10825-38. DOI: 10.1074/jbc.M512786200
Source: PubMed


One of the key pathological hallmarks of Alzheimer disease (AD) is the accumulation of paired helical filaments (PHFs) of hyperphosphorylated microtubule-associated protein Tau. Tandem mass spectrometry was employed to examine PHF-Tau post-translational modifications, in particular protein phosphorylation and ubiquitination, to shed light on their role in the early stages of Alzheimer disease. PHF-Tau from Alzheimer disease brain was affinity-purified by MC1 monoclonal antibody to isolate a soluble fraction of PHF-Tau in a conformation unique to human AD brain. A large number of phosphorylation sites were identified by employing a data-dependent neutral loss algorithm to trigger MS3 scans of phosphopeptides. It was found that soluble PHF-Tau is ubiquitinated at its microtubule-binding domain at residues Lys-254, Lys-311, and Lys-353, suggesting that ubiquitination of PHF-Tau may be an earlier pathological event than previously thought and that ubiquitination could play a regulatory role in modulating the integrity of microtubules during the course of AD. Tandem mass spectrometry data for ubiquitin itself indicate that PHF-Tau is modified by three polyubiquitin linkages, at Lys-6, Lys-11, and Lys-48. Relative quantitative analysis indicates that Lys-48-linked polyubiquitination is the primary form of polyubiquitination with a minor portion of ubiquitin linked at Lys-6 and Lys-11. Because modification by Lys-48-linked polyubiquitin chains is known to serve as the essential means of targeting proteins for degradation by the ubiquitin-proteasome system, and it has been reported that modification at Lys-6 inhibits ubiquitin-dependent protein degradation, a failure of the ubiquitin-proteasome system could play a role in initiating the formation of degradation-resistant PHF tangles.

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    • "Also, the large filamentary tangles might be toxic to neurons by acting as physical barriers in the cytoplasm. Therefore, neurons containing tau tangles actively activate diverse cell metabolisms to get rid of the abnormal protein aggregates from cytoplasm [14] [15]. This might be a great burden to a neuron that results in neuronal toxicity and neurodegeneration. "
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    ABSTRACT: Accumulation of abnormal tau aggregates in neuron is an important pathological signature in multiple neurodegenerative disorders including Alzheimer’s disease. Tau is a neuron specific microtubule-associated protein that regulates microtubule stability, which is critical for axonal outgrowth and synaptic plasticity. In a pathological condition, tau dissociates from microtubules and forms insoluble aggregates called neurofibrillary tangles (NFTs). The accumulation of NFTs in neuron directly correlates with microtubule dysfunction and neuronal degeneration. Due to the pathophysiological importance of tau, great efforts have been made to understand tau aggregation processes and find therapeutics to halt or reverse the processes. However, progress has been slow due to the lack of a suitable method for monitoring tau aggregation. In this mini-review, we will review the conventional methods for studying tau aggregation, and introduce recent cell-based sensor approaches that allow to monitoring tau aggregation in living cells.
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    • "Although tau acetylation has also been proposed to be pathogenic due to the removal of lysine residues available for ubiquitination (22), insufficient ubiquitination is unlikely to be the cause of tau accumulation in disease given that tau species present in NFTs are already polyubiquitinated (40). In fact, given that ubiquitination would also remove lysine residues available for acetylation, the finding that pathological tau species have been reported to be both ubiquitinated and hyperphosphorylated on KXGS motifs (19,40) would appear to provide additional evidence that decreased acetylation on these critical sites is involved in disease pathogenesis. "
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    ABSTRACT: The accumulation of hyperphosphorylated tau in neurofibrillary tangles (NFTs) is a neuropathological hallmark of tauopathies, including Alzheimer's disease (AD) and chronic traumatic encephalopathy, but effective therapies directly targeting the tau protein are currently lacking. Herein, we describe a novel mechanism in which the acetylation of tau on KXGS motifs inhibits phosphorylation on this same motif, and also prevents tau aggregation. Using a site-specific antibody to detect acetylation of KXGS motifs, we demonstrate that these sites are hypoacetylated in patients with AD, as well as a mouse model of tauopathy, suggesting that loss of acetylation on KXGS motifs renders tau vulnerable to pathogenic insults. Furthermore, we identify histone deacetylase 6 (HDAC6) as the enzyme responsible for the deacetylation of these residues, and provide proof of concept that acute treatment with a selective and blood–brain barrier-permeable HDAC6 inhibitor enhances acetylation and decreases phosphorylation on tau's KXGS motifs in vivo. As such, we have uncovered a novel therapeutic pathway that can be manipulated to block the formation of pathogenic tau species in disease.
    Full-text · Article · Aug 2013 · Human Molecular Genetics
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    • "As many as 30 different phosphorylation sites have been identified on tau [113], and although phosphorylation is the most comprehensively studied of the tau PTMs, filamentous tau is known to be extensively modified by several PTMs, including lysine-directed ubiquitylation [114]. We and others have identified at least three sites of ubiquitylation on tau [115] [116] [117]. Although most well known for its role in proteasomal degradation of proteins, ubiquitin is also the best known signal for endocytic sorting [118]. "
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