Chemical Inhibition of NAT10 Corrects Defects of Laminopathic Cells
Remodelin Nuclear Defects
Deregulation of A-type lamin proteins leads to disorganization of chromatin structure and misshapen nuclei, which are believed to underlie the pathologies of various human diseases, including the premature aging disorder Hutchinson Gilford progeria syndrome (HGPS) and various cancers. Larrieu et al. (p. 527) developed a small molecule, Remodelin, that not only improved nuclear shape defects of human lamin A/C–depleted cells, HGPS cells, and aged normal cells, but also decreased the levels of a DNA damage marker and improved global cellular fitness.
Abstract
Down-regulation and mutations of the nuclear-architecture proteins lamin A and C cause misshapen nuclei and altered chromatin organization associated with cancer and laminopathies, including the premature-aging disease Hutchinson-Gilford progeria syndrome (HGPS). Here, we identified the small molecule “Remodelin” that improved nuclear architecture, chromatin organization, and fitness of both human lamin A/C–depleted cells and HGPS-derived patient cells and decreased markers of DNA damage in these cells. Using a combination of chemical, cellular, and genetic approaches, we identified the acetyl-transferase protein NAT10 as the target of Remodelin that mediated nuclear shape rescue in laminopathic cells via microtubule reorganization. These findings provide insights into how NAT10 affects nuclear architecture and suggest alternative strategies for treating laminopathies and aging.
Get full access to this article
View all available purchase options and get full access to this article.
Already a Subscriber?Sign In
Supplementary Material
Summary
Materials and Methods
Figs S1 to S11
Movie S1
Resources
References and Notes
1
Worman H. J., Bonne G., “Laminopathies”: A wide spectrum of human diseases. Exp. Cell Res. 313, 2121–2133 (2007).
2
De Sandre-Giovannoli A., Bernard R., Cau P., Navarro C., Amiel J., Boccaccio I., Lyonnet S., Stewart C. L., Munnich A., Le Merrer M., Lévy N., Lamin a truncation in Hutchinson-Gilford progeria. Science 300, 2055 (2003).
3
Eriksson M., Brown W. T., Gordon L. B., Glynn M. W., Singer J., Scott L., Erdos M. R., Robbins C. M., Moses T. Y., Berglund P., Dutra A., Pak E., Durkin S., Csoka A. B., Boehnke M., Glover T. W., Collins F. S., Recurrent de novo point mutations in lamin A cause Hutchinson-Gilford progeria syndrome. Nature 423, 293–298 (2003).
4
Broers J. L., Raymond Y., Rot M. K., Kuijpers H., Wagenaar S. S., Ramaekers F. C., Nuclear A-type lamins are differentially expressed in human lung cancer subtypes. Am. J. Pathol. 143, 211–220 (1993).
5
Moss S. F., Krivosheyev V., de Souza A., Chin K., Gaetz H. P., Chaudhary N., Worman H. J., Holt P. R., Decreased and aberrant nuclear lamin expression in gastrointestinal tract neoplasms. Gut 45, 723–729 (1999).
6
Venables R. S., McLean S., Luny D., Moteleb E., Morley S., Quinlan R. A., Lane E. B., Hutchison C. J., Expression of individual lamins in basal cell carcinomas of the skin. Br. J. Cancer 84, 512–519 (2001).
7
Galiová G., Bártová E., Raska I., Krejcí J., Kozubek S., Chromatin changes induced by lamin A/C deficiency and the histone deacetylase inhibitor trichostatin A. Eur. J. Cell Biol. 87, 291–303 (2008).
8
Chen C. Y., Chi Y. H., Mutalif R. A., Starost M. F., Myers T. G., Anderson S. A., Stewart C. L., Jeang K. T., Accumulation of the inner nuclear envelope protein Sun1 is pathogenic in progeric and dystrophic laminopathies. Cell 149, 565–577 (2012).
9
Toth J. I., Yang S. H., Qiao X., Beigneux A. P., Gelb M. H., Moulson C. L., Miner J. H., Young S. G., Fong L. G., Blocking protein farnesyltransferase improves nuclear shape in fibroblasts from humans with progeroid syndromes. Proc. Natl. Acad. Sci. U.S.A. 102, 12873–12878 (2005).
10
Chimenti F., Bizzarri B., Maccioni E., Secci D., Bolasco A., Chimenti P., Fioravanti R., Granese A., Carradori S., Tosi F., Ballario P., Vernarecci S., Filetici P., A novel histone acetyltransferase inhibitor modulating Gcn5 network: Cyclopentylidene-[4-(4′-chlorophenyl)thiazol-2-yl)hydrazone. J. Med. Chem. 52, 530–536 (2009).
11
Chi Y. H., Haller K., Peloponese J. M., Jeang K. T., Histone acetyltransferase hALP and nuclear membrane protein hsSUN1 function in de-condensation of mitotic chromosomes. J. Biol. Chem. 282, 27447–27458 (2007).
12
Shen Q., Zheng X., McNutt M. A., Guang L., Sun Y., Wang J., Gong Y., Hou L., Zhang B., NAT10, a nucleolar protein, localizes to the midbody and regulates cytokinesis and acetylation of microtubules. Exp. Cell Res. 315, 1653–1667 (2009).
13
Li X., et al., J. Am. Chem. Soc. 134, 1982–1985 (2012).
14
Rodriguez R., Miller K. M., Forment J. V., Bradshaw C. R., Nikan M., Britton S., Oelschlaegel T., Xhemalce B., Balasubramanian S., Jackson S. P., Small-molecule-induced DNA damage identifies alternative DNA structures in human genes. Nat. Chem. Biol. 8, 301–310 (2012).
15
Chimnaronk S., Suzuki T., Manita T., Ikeuchi Y., Yao M., Suzuki T., Tanaka I., RNA helicase module in an acetyltransferase that modifies a specific tRNA anticodon. EMBO J. 28, 1362–1373 (2009).
16
Scaffidi P., Misteli T., Lamin A-dependent nuclear defects in human aging. Science 312, 1059–1063 (2006).
17
Capell B. C., Erdos M. R., Madigan J. P., Fiordalisi J. J., Varga R., Conneely K. N., Gordon L. B., Der C. J., Cox A. D., Collins F. S., Inhibiting farnesylation of progerin prevents the characteristic nuclear blebbing of Hutchinson-Gilford progeria syndrome. Proc. Natl. Acad. Sci. U.S.A. 102, 12879–12884 (2005).
18
Verstraeten V. L., Peckham L. A., Olive M., Capell B. C., Collins F. S., Nabel E. G., Young S. G., Fong L. G., Lammerding J., Protein farnesylation inhibitors cause donut-shaped cell nuclei attributable to a centrosome separation defect. Proc. Natl. Acad. Sci. U.S.A. 108, 4997–5002 (2011).
19
Glynn M. W., Glover T. W., Incomplete processing of mutant lamin A in Hutchinson-Gilford progeria leads to nuclear abnormalities, which are reversed by farnesyltransferase inhibition. Hum. Mol. Genet. 14, 2959–2969 (2005).
20
Wang Y., Ostlund C., Choi J. C., Swayne T. C., Gundersen G. G., Worman H. J., Blocking farnesylation of the prelamin A variant in Hutchinson-Gilford progeria syndrome alters the distribution of A-type lamins. Nucleus 3, 452–462 (2012).
21
Ibrahim M. X., Sayin V. I., Akula M. K., Liu M., Fong L. G., Young S. G., Bergo M. O., Targeting isoprenylcysteine methylation ameliorates disease in a mouse model of progeria. Science 340, 1330–1333 (2013).
22
Amin M. A., Matsunaga S., Ma N., Takata H., Yokoyama M., Uchiyama S., Fukui K., Fibrillarin, a nucleolar protein, is required for normal nuclear morphology and cellular growth in HeLa cells. Biochem. Biophys. Res. Commun. 360, 320–326 (2007).
23
Amin M. A., Matsunaga S., Uchiyama S., Fukui K., Depletion of nucleophosmin leads to distortion of nucleolar and nuclear structures in HeLa cells. Biochem. J. 415, 345–351 (2008).
24
Wang G., Gao X., Huang Y., Yao Z., Shi Q., Wu M., Nucleophosmin/B23 inhibits Eg5-mediated microtubule depolymerization by inactivating its ATPase activity. J. Biol. Chem. 285, 19060–19067 (2010).
25
Tamiello C., Kamps M. A., van den Wijngaard A., Verstraeten V. L., Baaijens F. P., Broers J. L., Bouten C. C., Soft substrates normalize nuclear morphology and prevent nuclear rupture in fibroblasts from a laminopathy patient with compound heterozygous LMNA mutations. Nucleus 4, 61–73 (2013).
26
Suzuki N., Del Villar K., Tamanoi F., Farnesyltransferase inhibitors induce dramatic morphological changes of KNRK cells that are blocked by microtubule interfering agents. Proc. Natl. Acad. Sci. U.S.A. 95, 10499–10504 (1998).
Information & Authors
Information
Published In
Science
Volume 344 | Issue 6183
2 May 2014
2 May 2014
Copyright
Copyright © 2014, American Association for the Advancement of Science.
Submission history
Received: 25 February 2014
Accepted: 4 April 2014
Published in print: 2 May 2014
Acknowledgments
We thank all members of the Jackson laboratory for help and support, J. Forment for discussions, and K. Dry and J. Travers for discussions and critical reading of the manuscript. We thank R. Belotserkovskaya for the GFP-H2B U2OS cell line and S. Heidorn and M. Garnett (Wellcome Trust Sanger Institute) for providing us with the cancer cells used in the lamin A/C expression screen. We thank Imagif and Institut de Chimie des Substances Naturelles, Centre National de la Recherche Scientifique (CNRS) de Gif-sur-Yvette, France, for proteomic analysis. Research in the Jackson laboratory is funded by Cancer Research UK program grant C6/A11224, the European Research Council, and the European Community Seventh Framework Programme grant agreement HEALTH-F2-2010-259893 (DDR). Core funding is provided by CRUK (C6946/A14492) and the Wellcome Trust (WT092096). S.P.J. receives his salary from the University of Cambridge, UK, supplemented by CRUK. D.L is funded by a European Molecular Biology Organization (EMBO) long-term fellowship ALTF 834-2011 and by a Project Grant from the Medical Research Council, UK MR/L019116/1. S.B. was funded by a EMBO long-term fellowship ALTF 93-2010 and CRUK. R.R. is supported by CNRS. M.D. is supported by the European Research Council grant DDREAM. A patent (application number 1405991.9) on the small molecule Remodelin and structural analogs for the treatment of disease has been filed by the University of Cambridge and CNRS. The data presented in this paper are included in the main paper and the supplementary materials. D.L. conceptualized the study and carried out all the experiments unless stated otherwise. S.B. cloned NAT10 expression constructs, performed the NAT10 structure analysis, produced high-resolution microscopy images, and contributed to live-cell imaging. R.R. designed and synthesized the small molecules and performed circular dichroism spectroscopy experiment. M.D. provided assistance with IF, fluorescence-activated cell sorting, and western blotting experiments. D.L. designed the experiments and analyzed the data. D.L. and S.P.J wrote the paper, with contributions from R.R., and S.B.
Authors
Metrics & Citations
Metrics
Article Usage
Altmetrics
Citations
Export citation
Select the format you want to export the citation of this publication.
Cited by
- Structural insights of human N-acetyltransferase 10 and identification of its potential novel inhibitors, Scientific Reports, 11, 1, (2021).https://doi.org/10.1038/s41598-021-84908-0
- Nuisance compounds in cellular assays, Cell Chemical Biology, 28, 3, (356-370), (2021).https://doi.org/10.1016/j.chembiol.2021.01.021
- Systematic screening identifies therapeutic antisense oligonucleotides for Hutchinson–Gilford progeria syndrome, Nature Medicine, 27, 3, (526-535), (2021).https://doi.org/10.1038/s41591-021-01262-4
- Arrow pushing in RNA modification sequencing, Chemical Society Reviews, (2021).https://doi.org/10.1039/D1CS00214G
- Interphase microtubules in nuclear organization and genome maintenance, Trends in Cell Biology, (2021).https://doi.org/10.1016/j.tcb.2021.03.014
- Quantitative nucleotide resolution profiling of RNA cytidine acetylation by ac4C-seq, Nature Protocols, 16, 4, (2286-2307), (2021).https://doi.org/10.1038/s41596-021-00501-9
- Evolution and diversification of the nuclear envelope, Nucleus, 12, 1, (21-41), (2021).https://doi.org/10.1080/19491034.2021.1874135
- NAT10 as a potential prognostic biomarker and therapeutic target for HNSCC, Cancer Cell International, 21, 1, (2021).https://doi.org/10.1186/s12935-021-02124-2
- A novel missense mutation of the NAT10 gene in a juvenile Schnauzer dog with chronic respiratory tract infections , Journal of Veterinary Internal Medicine, 35, 3, (1542-1546), (2021).https://doi.org/10.1111/jvim.16100
- The Keap1-Nrf2 System: A Mediator between Oxidative Stress and Aging, Oxidative Medicine and Cellular Longevity, 2021, (1-16), (2021).https://doi.org/10.1155/2021/6635460
- See more
Loading...
View Options
Get Access
Log in to view the full text
AAAS login provides access to Science for AAAS Members, and access to other journals in the Science family to users who have purchased individual subscriptions.
- Become a AAAS Member
- Activate your AAAS ID
- Purchase Access to Other Journals in the Science Family
- Account Help
Log in via OpenAthens.
Log in via Shibboleth.
More options
Register for free to read this article
As a service to the community, this article is available for free. Login or register for free to read this article.
Buy a single issue of Science for just $15 USD.
View options
PDF format
Download this article as a PDF file
Download PDF