2017 Guide of H3k27 methylation antibodies

 

1. Background information of H3K27me

In numerous organisms, methylation of lysine 27 on histone H3 (H3K27me) by Polycomb Repressive Complex 2 (PRC2) is a stamp of facultative heterochromatin. A Drosophila research identified and characterized the Polycomb group (PcG) protein complexes as writers and readers of H3K27me firstly and PcG maintains repression established in early development. The SET domain of the PRC2 component EZH2 requires both EED and SUZ12 to perform catalyzes H3K27me2/3; these proteins are the core components of PRC2. In the canonical Polycomb Repressive Complex 1 (PRC1), H3K27me3 can be recognized by a chromodomain protein. H2AK119 monoubiquitylation (H2Aub1) and chromatin compaction are thought as two ways to help mediate transcriptional repression for PRC1.

 

2. H3k27 methylation: H3k27me1, H3k27me2, H3k27me3 and H3k27 acetylation

Researches with mouse embryonic stem cell first indicate that perturbation of constitutive heterochromatin influences the distribution of H3K27me. Loss of H3K9me3 in pericentric heterochromatin in Suv39h double null cells was associated with new domains of H3K27me3 in patterns resembling those typical of H3K9me3 was showed in cytological observations.

Absence the EED subunit of PRC2 binding to H3K9me2 at the centromere can lead to the chromodomain protein normally binds H3K27me3 to restrict this histone modification to its normal location. H3K27me2/3 is redistributed to constitutive heterochromatin, including centromeres and reduced or lost at most of its normal locations when H3K9me3 was eliminated. Moreover, elimination of HP1 which is normally bound to H3K9me3 causes equivalent redistribution of H3K27me2/3 which will not influence H3K9me3, providing an evidence of cohabitation of these two marks on the same molecule.

Actively transcribed regions of the genome marked by H3K27me are generally distinct from those marked by 3H3K4me3 or H3K36me2/3. H3K4me3 and H3K27me3 can coexist in ‘‘bivalent’’ domains in ESCs, while they are mutually exclusive at HOX genes in Drosophila embryos and differentiated mammalian cells. In C. elegans embryos and murine mesenchymal progenitor cells, methylation of H3K27 at H3K36me region can be caused by loss of H3K36 methyltransferases. A human chondroblastomas mutation, H3.3K36 M, can cause global loss of H3K36me2/3 and induces increase of H3K27me3 in regions where lost H3K36me. Besides, in mouse and human cell lines, quantitative mass spectrometry experiments proved that H3K4me3 and H3K27me3 or H3K36me3 and H3K27me3 rarely co-exist on the same H3 molecule.

But there is more to H3K27 than methylation. Acetylation of H3K27 has also been reported. Since a lysine residue cannot be both methylated and acetylated you would expect H3K27ac to be antagonistic to the repression of gene expression by H3K27me2/3. Indeed, data are showing that H3K27ac is associated with active transcription and antagonism of H3K27me3 regulated genes.

Check 2017 ChIP Validated Histone H3k27me antibodies. Epicypher validated Abcam OEM supplier:

 
More details about the supplier

Point 1
Validated Original manufacturer.

ABclonal Inc. is one of the original manufactures which is famous for its Epigenetics antibodies such as H3K4me antibody and H3K9me antibody. It is the long-term OEM supplier for many top10 antibody companies and we also visited their factory for double confirmation.

Point 2
Lot number management

Lot number is unique for each batch of antibody and it can be taken as identity number of antibody.  ABclonal has effective batch management and it helps you maximize the reproducibility. We will share everything about the antibodies basing on Lot number.

Point 3
Antibody validation for each batch.

ABclonal is performing strict antibody validation for each batch. WB, IHC, IF, DB and ChIP are performed to make sure each Histone antibody has strong specificity and no cross-reactivity for multiple applications. It is the only one of 52 Histone antibody suppliers which passed ChIP test by EpiCypher in 2017. (Paper to be published)

3. Interplay between H3k27 methylation and other histone modifications.

 

PRC2-h3k27me antibody

Model of PRC2 responds to the chromatin environment to establish H3K27me domains.

Repressive features of constitutive heterochromatin including DNA methylation (5mC), H3K9me (K9me), and HP1 binding influence methylation of H3K27 by PRC2. Conversely, histone modifications associated with transcription such as H3K4me (K4me), H3K36me (K36me), and H3K27ac (K27ac), as well as RNA can directly inhibit PRC2 catalytic activity to prevent H3K27me at regions of active gene expression.

Check other 2017 validated Histone H3 product guide:


4. Related protein study of H3K27me

The H3K9me protein study has three parts, including writers, erasers and readers.
EZH1, EZH2, G9a and NSD3 are writers, UTX, JMJD and PHF8 are erasers and CBX2, CBX4, CBX6-8 and EED are readers.

 

Writers Find Antibodies introduction Reference
EZH1+EZH2

 EZH1 antibody;

EZH2 antibody

While PRC2-Ezh2 catalyzes H3K27me2/3 and its knockdown affects global H3K27me2/3 levels, PRC2-Ezh1 still weakly performs this function. In accordance, Ezh1 knockdown was ineffectual on global H3K27me2/3 levels. Margueron, Raphael; Li, Guohong; Sarma, Kavitha; Blais, Alexandre; Zavadil, Jiri; Woodcock, Christopher L.; Dynlacht, Brian D.; Reinberg, Danny. In Molecular Cell. 2008 32(4):503-518 Language: English. DOI: 10.1016/j.molcel.2008.11.004, Database: ScienceDirect
G9a  g9a (EHMT2 ) antibody Blockade of G9a can rescue H3K27me2 levels and neurodegeneration. Subbanna, S.; Nagre, N.N.; Shivakumar, M.; Umapathy, N.S.; Psychoyos, D.; Basavarajappa, B.S.. In Neuroscience. 31 January 2014 258:422-432 Language: English. DOI: 10.1016/j.neuroscience.2013.11.043, Database: ScienceDirect
NSD1, 2, 3  NSD1 antibody

NSD2 (WHSC1) antibody

NSD3 antibody

The nuclear receptor-binding SET domain (NSD) family, a part of the HMTase KMT3 family, is composed of three HMTases: NSD1, NSD2/MMSET/WHSC1, and NSD3/WHSC1L1 (hereafter NSD1, NSD2 and NSD3). The NSD proteins are oncogenes which highly expressed in numerous pathological conditions and are considered novel therapeutic targets in cancers. Morishita, Masayo; Mevius, Damiaan; di Luccio, Eric. In: BMC Structural Biology. Dec 12, 2014, Vol. 14, 25; BioMed Central Ltd. Language: English, Database: Academic OneFile
Erasers Find Antibodies introduction Reference
UTX The H3K27me3 demethylase UTX differentially performs in gender, it recurrently mutated in male T-ALL and escapes X-inactivation in female T-ALL blasts and normal T cells. The loss of Utx contributes to T-ALL formation in vivo and UTX inactivation confers sensitivity to H3K27me3 inhibition. Van der Meulen, J; Sanghvi, V; Mavrakis, K; Durinck, K; Fang, F; Matthijssens, F; Rondou, P; Rosen, M; Pieters, T; Vandenberghe, P; Delabesse, E; Lammens, T; De Moerloose, B; Menten, B; Van Roy, N; Verhasselt, B; Poppe, B; Benoit, Y; Taghon, T; Melnick, AM; Speleman, F; Wendel, HG; Van Vlierberghe, P. BLOOD; JAN 1, 2015; 125; 1; p13-p21, Database: Science Citation Index
JMJD  JMJD antibody The repressive H3K27me3, known to prevent chromatin remodelling and transcription, was removed from the SPINT2 but not the CCL18 gene locus under hypoxia or dimethyloxalylglycine-treatment. The H3K27me3 demethylase JMJD3 was required for CCL18 gene induction but not essential for induction of SPINT2. Dehne, Nathalie; Brüne, Bernhard. In BBA – Gene Regulatory Mechanisms. December 2016 1859(12):1490-1501 Language: English. DOI: 10.1016/j.bbagrm.2016.10.004, Database: ScienceDirect
PHF8  PHF8 antibody PHF8 regulates the levels of H3K9me2 and H3K27me2 in laryngeal and hypopharyngeal squamous cell carcinoma (LHSCC). Zhu, GC; Liu, LJ; She, L; Tan, HL; Wei, M; Chen, CH; Su, ZW; Huang, DH; Tian, YQ; Qiu, YZ; Liu, Y; Zhang, X. EPIGENOMICS; 2015; 7; 2; p143-p153, Database: Science Citation Index
Readers Find Antibodies introduction Reference
CBX2, 4, 6, 7, 8  CBX2 antibody

CBX4 antibody

CBX6 antibody

CBX8 antibody

The eight mammalian Cbx proteins are chromodomain-containing proteins involved in regulation of heterochromatin, gene expression, and developmental programs. They are evolutionarily related to the Drosophila HP1 (dHP1) and Pc (dPc) proteins that are core components of chromatin-associated complexes capable of recognizing repressive marks such as H3K9me3 and H3K27me3. However, the binding specificity and function of the human homologs, Cbx1–8, is still uncertainty which is currently in research. The five Pc homologs (Cbx2, -4, -6, -7, and -8) bind with lower affinity to H3K9me3 or H3K27me3 peptides and are unable to distinguish between these two marks. Liu, Xing-Yong; Zhang, Xian-Bo; Li, Ming-Hui; Zheng, Shu-Qing; Liu, Zhi-Long; Cheng, Yun-Ying; Wang, De-Shou. In Comparative Biochemistry and Physiology, Part B. January 2017 203:25-34 Language: English. DOI: 10.1016/j.cbpb.2016.09.001, Database: ScienceDirect
EED  EED antibody Loss of function somatic alterations of the Polycomb repressive complex 2 (PRC2) components (EED or SUZ12) in 92% of sporadic, 70% of NF1-associated and 90% of radiotherapyassociated MPNSTs. MPNSTs with PRC2 loss showed complete loss of H3K27me3 and aberrant transcriptional activation of multiple PRC2-repressed homeobox master regulators and their regulated developmental pathways. Lee, William; Qin, Li-Xuan; Singer, Samuel; Zheng, Deyou; Berger, Michael F; Chen, Yu; Chi, Ping; Teckie, Sewit; Wiesner, Thomas; Ran, Leili; Zhu, Sinan; Cao, Zhen; Liang, Yupu; Prieto Granada, Carlos N; Antonescu, Cristina R; Lin, Mingyan; Sboner, Andrea; Tap, William D; Fletcher, Jonathan A; Huberman, Kety H. Nature Genetics. Nov2014, Vol. 46 Issue 11, p1227-1232. 6p. DOI: 10.1038/ng.3095. , Database: Academic Search Complete

 

Reference

Tie F, Banerjee R, Stratton CA, et al. CBP-mediated acetylation of histone H3 lysine 27 antagonizes Drosophila Polycomb silencing. Development (Cambridge, England). 2009;136(18):3131-3141. doi:10.1242/dev.037127.

By Wiles, Elizabeth T; Selker, Eric U. In Genome architecture and expressionCurrent Opinion in Genetics & Development. April 2017 43:31-37 Language: English. DOI: 10.1016/j.gde.2016.11.001, Database: ScienceDirect