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H4K20 Monomethylation Faces the WNT

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H4K20 Monomethylation Faces the WNT COMMENTARY H4K20 monomethylation faces the WNT Gunnar Schotta1 Munich Center for Integrated Protein Science and Adolf Butenandt Institute, Ludwig Maximilians University, 80336 Munich, Germany rowth factor signaling pathways regulate a broad spectrum of G cellular processes ranging from proliferation to differentiation and tissue homeostasis. Activation of a signaling pathway ultimately leads to transcriptional changes in specific target genes. Although the molecular identities of many signaling pathway components have been revealed over the last years, there is still very little knowledge of how these components induce changes in the chromatin structure of target genes, a re- quirement for activation or repression of these genes. A new link is provided by an interesting paper in PNAS that demon- strates that in the context of Wnt signaling the histone methyltransferase SETD8 (PR-SET7, KMT5a, SET8) is recruited Fig. 1. Wnt signaling stimulates SETD8-mediated H4K20me1 at TCF/LEF binding sites (TBEs). (A)Inthe to enhancer regions of Wnt-regulated absence of Wnt ligand, cellular β-catenin is destabilized and cannot enter the nucleus. Wnt target genes genes. SETD8 establishes H4K20 mono- are constitutively bound by TCF/LEF transcription factors; however, transcription is blocked by binding of methylation (H4K20me1) at these regula- the repressor protein Groucho. (B) Under active Wnt signaling, β-catenin can enter the nucleus and tory regions, which is crucial for full displace Groucho from TCF/LEF. This allows for complex formation with the histone methyltransferase activation of these target genes (1). SETD8, which induces H4K20me1 at TBEs. Increased H4K20me1 is a prerequisite for full transcriptional The canonical Wnt signaling pathway activity of the Wnt target gene, possibly due to recruitment of currently unknown binding proteins. plays important roles for cellular differ- entiation during embryogenesis and tissue vation mark. On the one hand, there are tone modifications are thought to mediate homeostasis in adults. Dysregulation of correlative studies that link H4K20me1 effects on chromatin as interaction plat- this pathway is often found in cancer and with transcriptional activity (3, 4). On the forms for specific binding proteins, which other diseases. The Wnt-dependent acti- other hand, several publications demon- then mediate changes in the underlying vation of target genes is mediated through strate strong links of H4K20me1 with chromatin structure. Three binding pro- the action of T cell-specific transcription transcriptional repression and chromatin teins for H4K20me1 have been identified factor (TCF) and lymphoid enhancer- compaction (5–7). H4K20me1 is a very so far. N-CAPD3 and N-CAPG2 are sub- binding factor (LEF), which constitutively dynamic modification and could possibly units of the condensin II complex. Bind- bind to TCF binding elements (TBEs). In play different roles, depending on the cell- the absence of Wnt signaling, TCF/LEF cycle stage. Very low levels can be de- ing of these proteins to H4K20me1 in G2/M phase is important for mitotic cannot induce transcriptional activation tected in G1 and S phase, followed by a chromosome condensation (6). L3MBTL1 due to interaction with the corepressor massive increase in the G2/M transition Groucho. Under conditions of active Wnt (8). Previous analyses considered changes is a three malignant brain tumor (MBT) signaling, β-catenin is stabilized and enters only in bulk H4K20me1 levels. Cell cycle- domain protein and recognizes H4K20me1 the nucleus. It will then displace Groucho dependent changes at single loci have in the context of additional modifications from TCF/LEF and recruit other proteins, not been analyzed yet. (7). L3MBTL1 was shown to negatively leading to transcriptional activation of In metazoans, H4K20me1 is controlled regulate gene expression by inducing target genes (Fig. 1). Activation of Wnt by the histone methyltransferase SETD8. a compact chromatin structure (7, 9). It is target genes requires complex changes in Using a variety of knockdown and over- not clear whether any of these binding the underlying chromatin structure. In this expression approaches, Li et al. (1) dem- proteins is recruited to TBEs up- context, β-catenin was previously impli- onstrate that SETD8 is responsible for on Wnt signaling; however, due to their cated in the recruitment of chromatin re- H4K20me1 at human TBEs. Knockdown roles in gene repression and chromatin modelers and activating histone of SETD8 reduces H4K20me1 at these modifications to the c-myc gene (2). regulatory regions, and, consequently, condensation, this seems rather unlikely. To understand which histone mod- several of the tested Wnt target genes Therefore, mechanistic insight into the ac- ifications are implicated in the activation show reduced activation. Expression of tivating roles of H4K20me1 can be of Wnt target genes, Li et al. (1) probe 11 catalytically inactive SETD8 protein can- revealed only upon identification of other different histone methylation marks at not rescue these defects, and, therefore, binding proteins. the TBE of the AXIN2 gene in HEK293 H4K20me1 must directly play important cells. After stimulation of these cells with roles in activating transcription. This Wnt-conditioned medium, of all tested finding is difficult to explain, as currently Author contributions: G.S. wrote the paper. modifications, only H4K20me1 was sig- there is only limited knowledge about the The author declares no conflict of interest. nificantly increased. This is surprising, as mechanisms by which H4K20me1 can af- See companion article on page 3116. H4K20me1 is not a well-established acti- fect chromatin structure. In general, his- 1E-mail: [email protected]. www.pnas.org/cgi/doi/10.1073/pnas.1100147108 PNAS Early Edition | 1of2 Downloaded by guest on September 30, 2021 Li et al. (1) then go on to test whether In their PNAS paper, Li et al. (1) pro- TCF4 in cells that coexpress constitutively SETD8-mediated H4K20me1 regulates vide several lines of evidence that SETD8- active β-catenin, SETD8, and Groucho re- Wnt signaling in a more physiological set- mediated H4K20me1 at TBEs is impor- vealed that SETD8 can participate in the ting and decide to analyze zebrafish de- tant for full activation of Wnt target genes. TCF4 complex only when β-catenin is velopment. Morpholino knockdown of the But how is SETD8 recruited to TBEs? present and Groucho is displaced, suggest- SETD8 homolog Setd8a results in reduced To address this question, the authors test ing a novel mechanism for recruitment H4K20me1 at the Wnt8 target gene tbx6. whether SETD8 can interact with the of chromatin-modifying activities in Wnt Consistent with a Wnt-dependent recruit- signaling (Fig. 1). ment mechanism for Setd8a, H4K20me1 is The study by Li et al. (1) The study by Li et al. (1) identifies a reduced upon knockdown of Wnt8. Setd8a new function for H4K20me1 in transcrip- morphant embryos show developmental identifies a new function tional activation that will stimulate many defects, such as a shorter trunk and tail follow-up experiments. A central question region. These phenotypes mimic aspects of for H4K20me1 in will be the mechanism of transcriptional Wnt8 morphants; however, due to addi- activation. Is there a specific binding pro- tional functions of Setd8a outside of Wnt transcriptional activation. tein, and how does it contribute to open- signaling, the phenotypes are not expected ing the chromatin structure? Another to fully match. Notably, disruption of Setd8 TCF/LEF transcription factors. In a series question regards the inactivation of Wnt in the mouse results in a much stronger of interaction tests, they are able to dem- phenotype. Setd8 mouse mutant embryos signaling: Is there active removal of onstrate that SETD8 can form complexes H4K20me1 when Wnt target genes should die at the four-cell stage due to accumu- with TCF4 and LEF1. These complexes lation of DNA damage and mitotic defects be shut down? Two scenarios are possible. Drosophila exist only when Wnt signaling is active, and It might be that H4K20me1 is converted (10). Also, in , which is a phylo- it was therefore plausible to test whether genetically even more distant organism, to a higher methylation state by the action SETD8 is recruited through interaction of Suv4-20h enzymes (15). This could in- Setd8 has important functions in en- with β-catenin, as shown before for the duce a heterochromatin-like repressive state suring chromosome segregation and ge- Mll complex (2). However, no direct in- nome integrity (11, 12). Why is the de- teraction of SETD8 with β-catenin could around the TBE. Alternatively, H4K20me1 velopmental phenotype of zebrafish Setd8a be demonstrated. Therefore, Li et al. (1) could be actively removed by histone de- morphants comparably mild? One possi- test the intriguing hypothesis that the dis- methylases (6). An intriguing hypothesis is bility is that, in a knockdown situation, placement of Groucho by β-catenin could that such H4K20me1-antagonizing activities residual Setd8a activity could prevent ap- facilitate SETD8 binding. Groucho was could be targeted by Groucho complexes pearance of stronger phenotypes. Another shown before to interact with the high- that return to TBEs when β-catenin has left. explanation is that Setd8b, a second mobility group (HMG) domain of TCF4 SETD8 homolog in zebrafish, could have (14). The very same region is now mapped ACKNOWLEDGMENTS. Work in the G.S. labora- partially redundant functions (13). Future tory is supported by grants from the Deutsche by Li et al. (1) to bind SETD8. Therefore, Forschungsgemeinschaft, Bundesministerium für experiments are needed to distinguish Groucho might block the SETD8 binding Bildung und Forschung, and the Munich Center these possibilities. site in TCF4. Immunoprecipitation of for Integrated Protein Science. 1. Li Z, Nie F, Wang S, Li L (2011) Histone H4 Lys 20 mono- essential for mitotic entry and genomic stability. J Biol 11. Karachentsev D, Sarma K, Reinberg D, Steward R methylation by histone methylase SET8 mediates Wnt Chem 283:19478–19488. (2005) PR-Set7-dependent methylation of histone H4 target gene activation.
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