Tau Modification—Move Over Phosphate, Make Room for Acetylation

Before forming neurofibrillary tangles, tau may pick up a few acetyl groups along with the slew of phosphates it brings to the pathological inclusions in tauopathies such as Alzheimer’s disease and frontotemporal lobar dementia (FTLD). Researchers report in the March 22 issue of Nature Communications that tau is acetylated in vitro, in cell culture, in a mouse tauopathy model, and in several human tauopathies. They observed no acetylated tau in healthy tissue, suggesting acetylation could be part of the pathological process. The acetylated tau also made insoluble fibrils more rapidly than normal tau. Coupled with a report last fall that indicated acetylation slows degradation of tau, the findings suggest that this form of protein modification might play a major role in regulating tau toxicity.

First author Todd Cohen and senior author Virginia Lee led the study at the University of Pennsylvania School of Medicine in Philadelphia. The work confirms an earlier report of tau acetylation by Li Gan and colleagues at the Gladstone Institutes in San Francisco, California (see ARF related news story on Min et al., 2010). The current data are “very clear, black and white, very solid,” said Li-Huei Tsai of the Picower Institute in Cambridge, Massachusetts. “Now there are two papers indicating acetylation of tau in aggregation pathology…that is definitely reassuring,” said Tsai, who was not involved in either study.

Cohen joined Lee’s group two years ago with an interest in acetylation of proteins involved in neurodegeneration. He started by testing whether tau or α-synuclein could be acetylated in vitro. α-synuclein was not, but tau was. When he incubated recombinant tau protein with the acetyltransferase Creb-binding protein (CBP), the tau acquired acetyl groups. Tau in the presence of CBP also reacted with an antibody to acetylated lysines. In HEK293 human embryonic kidney cells, tau acetylation rose when Cohen added an inhibitor of deacetylation, trichostatin A. The researchers used mass spectrometry to determine that lysines 163, 280, 281, and 369 were modified under these conditions.

Lysines 280, 281, and 369 occur in the microtubule-binding regions of tau, and the authors hypothesized that the acetylation would interfere with the tau-microtubule interactions. In vitro, acetylated tau did not promote microtubule assembly as well as the un-acetylated form did. In cell culture, a mutant tau in which each of the four lysines was swapped for a glutamine—to mimic the acetylated state—bundled microtubules poorly in comparison to wild-type tau.

“At this point, we were fairly certain that tau, at least in these artificial situations—in vitro, in cells—was a substrate for acetyltransferases,” Lee said. But what about in a whole animal? To test this, the scientists engineered an antibody specific for the acetylated lysine 280 (K280) form of tau and used it to examine two mouse models of tauopathy: a PS19 line expressing the P301S tau mutant, and one expressing that tau mutation plus the V717F human amyloid precursor protein mutant (Hurtado et al., 2010). Both evinced immunoreactivity with the acetyl-tau antibody in the cortex and hippocampus, compared to wild-type mice. The double-mutant mice showed greater immunoreactivity as they aged. The acetyl-tau signal frequently co-localized with that for the AT8 phospho-tau antibody and with thioflavin S staining for amyloid, indicating that tau tangles contain acetylated, phosphorylated tau.

Finally, the researchers examined tau acetylation in human samples, tapping coauthor John Trojanowski’s extensive set of cortical sections from people who died from various tauopathies. While they observed no signal from the acetyl-tau antibody in control samples, they did see antibody staining in Alzheimer’s disease, frontotemporal lobar dementia with tau pathology (both corticobasal degeneration and progressive supranuclear palsy), argyrophilic grain disease, tangle predominant senile dementia, and even some signal from Guam parkinsonism-dementia complex. The only negative result was from Pick’s disease, in which the predominant tau isoform lacks K280. Though sample numbers were small, it appears tau acetylation may be a common feature of many tauopathies.

Lee suggested that as such, tau acetylation might be useful as a biomarker to distinguish different forms of frontotemporal lobar dementia (FTLD). While some FTLDs have tau pathology, others exhibit inclusions of the protein TDP-43 instead, but levels of phosphorylated tau are the same between the two groups. Lee speculated that acetyl-tau levels in the cerebrospinal fluid might allow clinicians to differentiate patients that need tau-based therapies from those that would be best served by TDP-43-related drugs. No such drugs are currently approved, and, at this point, the idea is only “a hope and a prayer,” Lee said, but she thinks it is a possibility worth exploring.

“What is the significance of this post-translational modification of tau in Alzheimer’s disease pathogenesis?” asked Khalid Iqbal of the New York State Institute for Basic Research in Staten Island, who was not involved in the study. Acetylation might promote phosphorylation, or block tau degradation, as suggested in Gan’s study. Acetylated tau is also more fibrillogenic, which could be a key pathological change. Using sedimentation analysis, electron microscopy, and thioflavin T staining to examine tau aggregation, Lee and colleagues found that acetylated tau formed fibrils much faster than non-acetylated. However, the current evidence is not yet enough to convince Iqbal that acetylation stands among the most important factors in disease. For example, he noted, it is not yet known how much of tau in fibrils is actually acetylated.

A key step toward understanding the importance of acetylation will be to identify the enzymes that acetylate and deacetylate tau. These enzymes would also be potential therapeutic targets, Lee noted. Cohen’s experiments with histone deacetylase 6 suggest it affects tau acetylation, while Gan’s study pointed to roles for histone acetyltransferase p300 and the deacetylase SIRT1; none has yet been proven to be a primary modifier of tau in vivo. Perhaps both deacetylases are correct, Tsai suggested: “I would not be surprised if there were redundant functions.”

Also unknown at this point is whether tau acetylation is a purely pathological phenomenon, or whether is serves some function in a healthy cell. One of the acetylation motifs in tau also appears in the related microtubule associated proteins MAP2 and MAP4. The study authors suggest that perhaps acetylation is a mechanism to regulate MAP-microtubule binding. Cohen might not have been able to observe acetylation in normal samples, Lee suggested, because deacetylation happens quickly. “I think it is likely that there is a role” in normal development, said Mark Mattson of the National Institute on Aging in Baltimore, Maryland. He speculated, for example, that acetylation might act to knock tau off microtubules when axons reach the appropriate length and stop growing.—Amber Dance

Comments

  1. From a single human gene, many different forms of tau protein could arise. Some of these forms come from alternative splicing of the nuclear RNA transcript, but other forms are the consequence of a post-translational modification—phosphorylation. Recently, an additional modification for tau protein has been indicated—acetylation (Min et al., 2010).

    In the last issue of Nature Communications, the group of Virginia Lee has observed that tau acetylation takes place in a key residue, lysine 280, which plays a role in the interaction of tau with microtubules. This finding shows how acetylation could regulate one of the main tau functions—its interaction with microtubules—but the report also opens the door for future experiments. Cohen et al. reported that tau deacetylation could occur through histone deacetylase 6 (HDA6), a deacetylase that is inhibited by tau. This hints that tau protein could self-regulate its deacetylation. This could be tested in the future. Secondly, it remains to be determined if acetylated tau could be deacetylated by more than one deacetylase. Lastly, as indicated in this paper, tau-lysine 280 is present in a motif that is similar to those present in other microtubule-associated proteins like MAP2 or MAP4. Thus, acetylation could be a common mechanism to regulate the interaction of some MAPs with microtubules, as the authors have proposed.

    In summary, this is an interesting and seminal work on the regulation of tau function.

    References:

    Min SW, Cho SH, Zhou Y, Schroeder S, Haroutunian V, Seeley WW, Huang EJ, Shen Y, Masliah E, Mukherjee C, Meyers D, Cole PA, Ott M, Gan L. Acetylation of tau inhibits its degradation and contributes to tauopathy. Neuron. 2010 Sep 23;67(6):953-66. PubMed.

    View all comments by Jesus Avila

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References

News Citations

  1. Tau Timing: New Findings on Disease Progression, Clearance

Paper Citations

  1. Min SW, Cho SH, Zhou Y, Schroeder S, Haroutunian V, Seeley WW, Huang EJ, Shen Y, Masliah E, Mukherjee C, Meyers D, Cole PA, Ott M, Gan L. Acetylation of tau inhibits its degradation and contributes to tauopathy. Neuron. 2010 Sep 23;67(6):953-66. PubMed.
  2. Hurtado DE, Molina-Porcel L, Iba M, Aboagye AK, Paul SM, Trojanowski JQ, Lee VM. A{beta} accelerates the spatiotemporal progression of tau pathology and augments tau amyloidosis in an Alzheimer mouse model. Am J Pathol. 2010 Oct;177(4):1977-88. PubMed.

Other Citations

  1. PS19 line

Further Reading

Papers

  1. Liu F, Iqbal K, Grundke-Iqbal I, Hart GW, Gong CX. O-GlcNAcylation regulates phosphorylation of tau: a mechanism involved in Alzheimer's disease. Proc Natl Acad Sci U S A. 2004 Jul 20;101(29):10804-9. PubMed.
  2. Liu F, Shi J, Tanimukai H, Gu J, Grundke-Iqbal I, Iqbal K, Gong CX. Reduced O-GlcNAcylation links lower brain glucose metabolism and tau pathology in Alzheimer's disease. Brain. 2009 Jul;132(Pt 7):1820-32. PubMed.
  3. Inouye H, Sharma D, Goux WJ, Kirschner DA. Structure of core domain of fibril-forming PHF/Tau fragments. Biophys J. 2006 Mar 1;90(5):1774-89. PubMed.
  4. Zambrano CA, Egaña JT, Núñez MT, Maccioni RB, González-Billault C. Oxidative stress promotes tau dephosphorylation in neuronal cells: the roles of cdk5 and PP1. Free Radic Biol Med. 2004 Jun 1;36(11):1393-402. PubMed.

Primary Papers

  1. Cohen TJ, Guo JL, Hurtado DE, Kwong LK, Mills IP, Trojanowski JQ, Lee VM. The acetylation of tau inhibits its function and promotes pathological tau aggregation. Nat Commun. 2011;2:252. PubMed.

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