Abstract
Clipping of the histone H3 N-terminal tail has been implicated in multiple fundamental biological processes for a growing list of eukaryotes. H3 clipping, serving as an irreversible process to permanently remove some post-translational modifications (PTMs), may lead to noticeable changes in chromatin dynamics or gene expression. The eukaryotic model organism Tetrahymena thermophila is among the first few eukaryotes that exhibits H3 clipping activity, wherein the first six amino acids of H3 are cleaved off during vegetative growth. Clipping only occurs in the transcriptionally silent micronucleus of the binucleated T. thermophila, thus offering a unique opportunity to reveal the role of H3 clipping in epigenetic regulation. However, the physiological functions of the truncated H3 and its protease(s) for clipping remain elusive. Here, we review the major findings of H3 clipping in T. thermophila and highlight its association with histone modifications and cell cycle regulation. We also summarize the functions and mechanisms of H3 clipping in other eukaryotes, focusing on the high diversity in terms of protease families and cleavage sites. Finally, we predict several protease candidates in T. thermophila and provide insights for future studies.
This is a preview of subscription content, access via your institution.




Data availability
Most datasets generated or analyzed during this study are included in this article (and its supplementary material), and the unpublished data in this article is available from the corresponding author upon reasonable request.
References
Adams-Cioaba MA, Krupa JC, Xu C, Mort JS, Min J (2011) Structural basis for the recognition and cleavage of histone H3 by cathepsin L. Nat Commun 2:197
Ali MAM, Garcia-Vilas JA, Cromwell CR, Hubbard BP, Hendzel MJ, Schulz R (2021) Matrix metalloproteinase-2 mediates ribosomal RNA transcription by cleaving nucleolar histones. FEBS J 288:6736–6751
Allis CD, Gorovsky MA (1981) Histone phosphorylation in macro- and micronuclei of Tetrahymena thermophila. Biochemistry 20:3828–3833
Allis CD, Wiggins JC (1984) Proteolytic processing of micronuclear H3 and histone phosphorylation during conjugation in Tetrahymena thermophila. Exp Cell Res 153:287–298
Allis CD, Glover CVC, Gorovsky MA (1979) Micronuclei of Tetrahymena contain two types of histone H3. Proc Natl Acad Sci USA 76:4857–4861
Allis CD, Bowen JK, Abraham GN, Glover CVC, Gorovsky MA (1980) Proteolytic processing of histone H3 in chromatin: a physiologically regulated event in Tetrahymena micronuclei. Cell 20:55–64
Allis CD, Allen RL, Wiggins JC, Chicoine LG, Richman R (1984) Proteolytic processing of H1-like histones in chromatin: a physiologically and developmentally regulated event in Tetrahymena micronuclei. J Cell Biol 99:1669–1677
Azad GK, Tomar RS (2014) Proteolytic clipping of histone tails: the emerging role of histone proteases in regulation of various biological processes. Mol Biol Rep 41:2717–2730
Chalker DL, Meyer E, Mochizuki K (2013) Epigenetics of ciliates. Cold Spring Harb Perspect Biol 5:a017764
Chapman HA, Riese RJ, Shi GP (1997) Emerging roles for cysteine proteases in human biology. Annu Rev Physiol 59:63–88
Cheng T, Wang Y, Huang J, Chen X, Zhao X, Gao S, Song W (2019) Our recent progress in epigenetic research using the model ciliate, Tetrahymena thermophila. Mar Life Sci Technol 1:4–14
Cheung P, Schaffert S, Chang SE, Dvorak M, Donato M, Macaubas C, Foecke MH, Li T-M, Zhang L, Coan JP, Schulert GS, Grom AA, Henderson LA, Nigrovic PA, Elias JE, Gozani O, Mellins ED, Khatri P, Utz PJ, Kuo AJ (2021) Repression of CTSG, ELANE and PRTN3-mediated histone H3 proteolytic cleavage promotes monocyte-to-macrophage differentiation. Nat Immunol 22:711–722
Daura E, Tegelberg S, Yoshihara M, Jackson C, Simonetti F, Aksentjeff K, Ezer S, Hakala P, Katayama S, Kere J, Lehesjoki A-E, Joensuu T (2021) Cystatin B-deficiency triggers ectopic histone H3 tail cleavage during neurogenesis. Neurobiol Dis 156:105418
Dhaenens M (2021) Histone clipping: the punctuation in the histone code. EMBO Rep 22:e53440
Dhaenens M, Glibert P, Meert P, Vossaert L, Deforce D (2015) Histone proteolysis: a proposal for categorization into ‘clipping’ and ‘degradation.’ BioEssays 37:70–79
Duan L, Cheng T, Wei F, Qiao Y, Wang C, Warren A, Niu J, Wang Y (2021) New contribution to epigenetic studies: isolation of micronuclei with high purity and DNA integrity in the model ciliated protist Tetrahymena thermophila. Eur J Protistol 80:125804
Duarte LF, Young ARJ, Wang Z, Wu H-A, Panda T, Kou Y, Kapoor A, Hasson D, Mills NR, Ma’ayan A, Narita M, Bernstein E (2014) Histone H3.3 and its proteolytically processed form drive a cellular senescence programme. Nat Commun 5:5210
Duncan EM, Muratore-Schroeder TL, Cook RG, Garcia BA, Shabanowitz J, Hunt DF, Allis CD (2008) Cathepsin L proteolytically processes histone H3 during mouse embryonic stem cell differentiation. Cell 135:284–294
Edgar RC (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 32:1792–1797
Falk MM, Grigera PR, Bergmann IE, Zibert A, Multhaup G, Beck E (1990) Foot-and-mouth disease virus protease 3C induces specific proteolytic cleavage of host cell histone H3. J Virol 64:748–756
Felsenfeld G (2014) A brief history of epigenetics. Cold Spring Harb Perspect Biol 6:a018200
Ferrari KJ, Amato S, Noberini R, Toscani C, Fernandez-Perez D, Rossi A, Conforti P, Zanotti M, Bonaldi T, Tamburri S, Pasini D (2021) Intestinal differentiation involves cleavage of histone H3 N-terminal tails by multiple proteases. Nucleic Acids Res 49:791–804
Guex N, Peitsch MC, Schwede T (2009) Automated comparative protein structure modeling with SWISS-MODEL and Swiss-PdbViewer: a historical perspective. Electrophoresis 30:S162–S173
Herrera-Solorio AM, Vembar SS, MacPherson CR, Lozano-Amado D, Meza GR, Xoconostle-Cazares B, Martins RM, Chen P, Vargas M, Scherf A, Hernandez-Rivas R (2019) Clipped histone H3 is integrated into nucleosomes of DNA replication genes in the human malaria parasite Plasmodium falciparum. EMBO Rep 20:e46331
Howe CG, Gamble MV (2015) Enzymatic cleavage of histone H3: a new consideration when measuring histone modifications in human samples. Clin Epigenet 7:7
Jiang L, Wang C, Zhuang W, Li S, Hu X (2021) Taxonomy, phylogeny, and geographical distribution of the little-known Helicoprorodon multinucleatum Dragesco, 1960 (Ciliophora, Haptorida) and key to species within the genus. Eur J Protistol 78:125769
Karrer KM (2012) Nuclear dualism. Methods Cell Biol 109:29–52
Khalkhali-Ellis Z, Goossens W, Margaryan NV, Hendrix MJC (2014) Cleavage of histone 3 by Cathepsin D in the involuting mammary gland. PLoS ONE 9:e103230
Kim K, Punj V, Kim J-M, Lee S, Ulmer TS, Lu W, Rice JC, An W (2016) MMP-9 facilitates selective proteolysis of the histone H3 tail at genes necessary for proficient osteoclastogenesis. Genes Dev 30:208–219
Kim K, Shin Y, Kim J, Ulmer TS, An W (2018) H3K27me1 is essential for MMP-9-dependent H3N-terminal tail proteolysis during osteoclastogenesis. Epigenet Chromatin 11:23
Kumar S, Stecher G, Tamura K (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33:1870–1874
Li Y, Chen X, Wu K, Pan J, Long H, Yan Y (2020) Characterization of simple sequence repeats (SSRs) in ciliated protists inferred by comparative genomics. Microorganisms 8:662
Lin R, Cook RG, Allis CD (1991) Proteolytic removal of core histone amino termini and dephosphorylation of histone H1 correlate with the formation of condensed chromatin and transcriptional silencing during Tetrahymena macronuclear development. Genes Dev 5:1601–1610
Liu W, Xie Y, Ma J, Luo X, Nie P, Zuo Z, Lahrmann U, Zhao Q, Zheng Y, Zhao Y, Xue Y, Ren J (2015) IBS: an illustrator for the presentation and visualization of biological sequences. Bioinformatics 31:3359–3361
Liu H, Wang C, Lee S, Deng Y, Wither M, Oh S, Ning F, Dege C, Zhang Q, Liu X, Johnson AM, Zang J, Chen Z, Janknecht R, Hansen K, Marrack P, Li C-Y, Kappler JW, Hagman J, Zhang G (2017) Clipping of arginine-methylated histone tails by JMJD5 and JMJD7. Proc Natl Acad Sci USA 114:E7717–E7726
Liu H, Hu M, Wang Q, Cheng L, Zhang Z (2018a) Role of papain-like cysteine proteases in plant development. Front Plant Sci 9:1717
Liu H, Wang C, Lee S, Ning F, Wang Y, Zhang Q, Chen Z, Zang J, Nix J, Dai S, Marrack P, Hagman J, Kappler J, Zhang G (2018b) Specific recognition of arginine methylated histone tails by JMJD5 and JMJD7. Sci Rep 8:3275
Liu Y, Nan B, Niu J, Kapler GM, Gao S (2021) An optimized and versatile counter-flow centrifugal elutriation workflow to obtain synchronized eukaryotic cells. Front Cell Dev Biol 9:905–917
Lu S, Wang J, Chitsaz F, Derbyshire MK, Geer RC, Gonzales NR, Gwadz M, Hurwitz DI, Marchler GH, Song JS, Thanki N, Yamashita RA, Yang M, Zhang D, Zheng C, Lanczycki CJ, Marchler-Bauer A (2020) CDD/SPARCLE: the conserved domain database in 2020. Nucleic Acids Res 48:D265–D268
Mahendra G, Kanungo MS (2000) Age-related and steroid induced changes in the histones of the quail liver. Arch Gerontol Geriatr 30:109–114
Mandal P, Azad GK, Tomar RS (2012) Identification of a novel histone H3 specific protease activity in nuclei of chicken liver. Biochem Biophys Res Commun 421:261–267
Mandal P, Verma N, Chauhan S, Tomar RS (2013) Unexpected histone H3 tail-clipping activity of glutamate dehydrogenase. J Biol Chem 288:18743–18757
Martindale DW, Allis CD, Bruns PJ (1982) Conjugation in Tetrahymena thermophila: a temporal analysis of cytological stages. Exp Cell Res 140:227–236
Melo FR, Vita F, Berent-Maoz B, Levi-Schaffer F, Zabucchi G, Pejler G (2014) Proteolytic histone modification by mast cell tryptase, a serglycin proteoglycan-dependent secretory granule protease. J Biol Chem 289:7682–7690
Melo FR, Wallerman O, Paivandy A, Calounova G, Gustafson A-M, Sabari BR, Zabucchi G, Allis CD, Pejler G (2017) Tryptase-catalyzed core histone truncation: a novel epigenetic regulatory mechanism in mast cells. J Allergy Clin Immunol 140:474–485
Miao W, Xiong J, Bowen J, Wang W, Liu Y, Braguinets O, Grigull J, Pearlman RE, Orias E, Gorovsky MA (2009) Microarray analyses of gene expression during the Tetrahymena thermophila life cycle. PLoS ONE 4:e4429
Mishra RN, Kanungo MS (1994) Alterations in histones of the liver and oviduct of Japanese quail during aging. Mol Biol Rep 20:15–18
Mochizuki K, Fine NA, Fujisawa T, Gorovsky MA (2002) Analysis of a piwi-related gene implicates small RNAs in genome rearrangement in Tetrahymena. Cell 110:689–699
Mort JS, Buttle DJ (1997) Cathepsin B. Int J Biochem Cell Biol 29:715–720
Musil D, Zucic D, Turk D, Engh RA, Mayr I, Huber R, Popovic T, Turk V, Towatari T, Katunuma N, Bode W (1991) The refined 2.15 Å X-ray crystal structure of human liver cathepsin B: the structural basis for its specificity. EMBO J 10:2321–2330
Orias E (2000) Toward sequencing the Tetrahymena genome: exploiting the gift of nuclear dimorphism. J Eukaryot Microbiol 47:328–333
Orias E, Cervantes MD, Hamilton EP (2011) Tetrahymena thermophila, a unicellular eukaryote with separate germline and somatic genomes. Res Microbiol 162:578–586
Papazyan R, Voronina E, Chapman JR, Luperchio TR, Gilbert TM, Meier E, Mackintosh SG, Shabanowitz J, Tackett AJ, Reddy KL, Coyne RS, Hunt DF, Liu Y, Taverna SD (2014) Methylation of histone H3K23 blocks DNA damage in pericentric heterochromatin during meiosis. Elife 3:e02996
Price MN, Dehal PS, Arkin AP (2010) FastTree 2-approximately maximum-likelihood trees for large alignments. PLoS ONE 5:e9490
Qiao J, Xu J, Bo T, Wang W (2017) Micronucleus-specific histone H1 is required for micronuclear chromosome integrity in Tetrahymena thermophila. PLoS ONE 12:e0187475
Rice JC, Weekley BH, Kanholm T, Chen Z, Lee S, Fernandez DJ, Abrahamson R, Castaldi A, Borok Z, Dynlacht BD, An W (2021) MMP-2 is a novel histone H3 N-terminal protease necessary for myogenic gene activation. Epigenet Chromatin 14:23
Santos-Rosa H, Kirmizis A, Nelson C, Bartke T, Saksouk N, Cote J, Kouzarides T (2009) Histone H3 tail clipping regulates gene expression. Nat Struct Mol Biol 16:17–22
Shen J, Xiang X, Chen L, Wang H, Wu L, Sun Y, Ma L, Gu X, Liu H, Wang L, Yu Y-N, Shao J, Huang C, Chin YE (2017) JMJD5 cleaves monomethylated histone H3 N-tail under DNA damaging stress. EMBO Rep 18:2131–2143
Sheng Y, Duan L, Cheng T, Qiao Y, Stover NA, Gao S (2020) The completed macronuclear genome of a model ciliate Tetrahymena thermophila and its application in genome scrambling and copy number analyses. Sci China Life Sci 63:1534–1542
Sheng Y, Pan B, Wei F, Wang Y, Gao S (2021) Case study of the response of N6-methyladenine DNA modification to environmental stressors in the unicellular eukaryote Tetrahymena thermophila. mSphere 6:e01208-01220
Shin Y, Kim S, Ghate NB, Rhie SK, An W (2022) MMP-9 drives the melanomagenic transcription program through histone H3 tail proteolysis. Oncogene 41:560–570
Stillman B (2018) Histone modifications: insights into their influence on gene expression. Cell 175:6–9
Sweet MT, Allis CD (2006) Induction of conjugation in Tetrahymena. Cold Spring Harb Protoc 2006:pdb.prot4499
Tan M, Luo H, Lee S, Jin F, Yang JS, Montellier E, Buchou T, Cheng Z, Rousseaux S, Rajagopal N, Lu Z, Ye Z, Zhu Q, Wysocka J, Ye Y, Khochbin S, Ren B, Zhao Y (2011) Identification of 67 histone marks and histone lysine crotonylation as a new type of histone modification. Cell 146:1016–1028
Taverna SD, Ueberheide BM, Liu Y, Tackett AJ, Diaz RL, Shabanowitz J, Chait BT, Hunt DF, Allis CD (2007) Long-distance combinatorial linkage between methylation and acetylation on histone H3 N termini. Proc Natl Acad Sci USA 104:2086–2091
Turk B, Turk V, Turk D (1996) Structural and functional aspects of papain-like cysteine proteinases and their protein inhibitors. Biol Chem 378:141–150
Vaughan RM, Kupai A, Rothbart SB (2021) Chromatin regulation through ubiquitin and ubiquitin-like histone modifications. Trends Biochem Sci 46:258–269
Villar-Garea A, Imhof A (2006) The analysis of histone modifications. Biochim Biophys Acta Proteins Proteom 1764:1932–1939
Vossaert L, Meert P, Scheerlinck E, Glibert P, Van Roy N, Heindryckx B, De Sutter P, Dhaenens M, Deforce D (2014) Identification of histone H3 clipping activity in human embryonic stem cells. Stem Cell Res 13:123–134
Waterhouse A, Bertoni M, Bienert S, Studer G, Tauriello G, Gumienny R, Heer FT, de Beer TAP, Rempfer C, Bordoli L, Lepore R, Schwede T (2018) SWISS-MODEL: homology modelling of protein structures and complexes. Nucleic Acids Res 46:W296–W303
Wu M, Allis CD, Sweet MT, Cook RG, Thatcher TH, Gorovsky MA (1994) Four distinct and unusual linker proteins in a mitotically dividing nucleus are derived from a 71-kilodalton polyprotein, lack p34cdc2 sites, and contain protein kinase A sites. Mol Cell Biol 14:10–20
Xiong J, Lu Y, Feng J, Yuan D, Tian M, Chang Y, Fu C, Wang G, Zeng H, Miao W (2013) Tetrahymena functional genomics database (TetraFGD): an integrated resource for Tetrahymena functional genomics. Database (oxford) 2013:bat008
Xu J, Zhao X, Mao F, Basrur V, Ueberheide B, Chait BT, Allis CD, Taverna SD, Gao S, Wang W, Liu Y (2021) A Polycomb repressive complex is required for RNAi-mediated heterochromatin formation and dynamic distribution of nuclear bodies. Nucleic Acids Res 49:5407–5425
Xue Y, Vashisht AA, Tan Y, Su T, Wohlschlegel JA (2014) PRB1 is required for clipping of the histone H3 N terminal tail in Saccharomyces cerevisiae. PLoS ONE 9:e90496
Yi SJ, Kim K (2018) Histone tail cleavage as a novel epigenetic regulatory mechanism for gene expression. BMB Rep 51:211–218
Zhang T, Li C, Zhang X, Wang C, Roger AJ, Gao F (2021) Characterization and comparative analyses of mitochondrial genomes in single-celled eukaryotes to shed light on the diversity and evolution of linear molecular architecture. Int J Mol Sci 22:2546
Zhao X, Li Y, Duan L, Chen X, Mao F, Juma M, Liu Y, Song W, Gao S (2020) Functional analysis of the methyltransferase SMYD in the single-cell model organism Tetrahymena thermophila. Mar Life Sci Technol 2:109–122
Zhao L, Gao F, Gao S, Liang Y, Long H, Lv Z, Su Y, Ye N, Zhang L, Zhao C, Wang X, Song W, Zhang S, Dong B (2021) Biodiversity-based development and evolution: the emerging research systems in model and non-model organisms. Sci China Life Sci 64:1236–1280
Zhou P, Wu E, Alam HB, Li Y (2014) Histone cleavage as a mechanism for epigenetic regulation: current insights and perspectives. Curr Mol Med 14:1164–1172
Acknowledgements
This work is supported by the National Natural Science Foundation of China (32125006, 32070437), China National Postdoctoral Program for Innovative Talents (BX2021277), China Postdoctoral Science Foundation (2021M690144), Natural Science Foundation of Shandong Province of China (ZR2021QC046), and Postdoctoral Applied Research Project of Qingdao. We appreciate the computing resources provided on IEMB-1, a high-performance computing cluster operated by the Institute of Evolution and Marine Biodiversity. We are grateful to Dr. Weibo Song (Ocean University of China, OUC) and Dr. Borong Lu (OUC) for their valuable inputs and suggestions to improve the manuscript.
Author information
Authors and Affiliations
Contributions
FW and BP collected the data and wrote the paper. JHD revised the article. YYW, YLS and SG designed, directed and revised this review. All the authors read and approved the final manuscript.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Animal and human rights statement
This article does not contain human participates or animals.
Additional information
Edited by Jiamei Li.
Special Topic: Ciliatology.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Wei, F., Pan, B., Diao, J. et al. The micronuclear histone H3 clipping in the unicellular eukaryote Tetrahymena thermophila. Mar Life Sci Technol 4, 584–594 (2022). https://doi.org/10.1007/s42995-022-00151-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s42995-022-00151-0
Keywords
- Histone H3 tail
- H3 clipping protease
- Histone post-translational modifications (PTMs)
- Cell development