2017 Guide of H3k79 methylation antibodies

 

1. Function of H3k79 methylation

1.1 Transcriptional regulation

H3K79 methylation levels show strong correlation with transcriptional activity.
H3K79 methylation may be important for the recruitment of transcription machinery or it might act as a landing platform for various transcription activators or it may inhibit the binding of transcription repressors. Methylation of H3K79 has also been implicated in the reactivation of tumor suppressor genes upon DNA demethylation.

1.2 Telomeric silencing

Cells overexpressing Dot1 show detectable H3K79 methylation spreading into silent regions of the chromatin. The presence of H3K79 methylation is considered to displace Sir proteins from silent chromatin, causing silencing defects.

1.3 Cell Cycle

Histone modifications such as H3K79 methylation have recently been proven to be crucial contributors to cell-cycle regulation.
In S. cerevisiae, the levels of H3K79me3 remain unchanged throughout the cell cycle, whereas the H3K79me2 levels increase gradually from the G1 to S phase and then further in the G2/M phase

1.4 DNA repair

The first evidence was binding of the tandem tudor domain (a chromodomain) of human 53BP1 (ortholog of Rad9 in yeast) protein to methylated H3K79 and its recruitment to DNA DSBs. The mutation of histone H3 lysine 79 or depletion of hDOT1L inhibited the recruitment of 53BP1 to DSBs as H3K79 methylation is impaired in both cases.

 

2. Three types of H3k79 methylation: H3k79me1, H3k79me2, H3k79me3

In Drosophila, chromatin immunoprecipitation (ChIP) coupled with gene expression microarray (ChIP–chip) revealed that histone acetylation, H3K4me2, H3K4me3, and H3K79me2 positively correlated with active gene transcription (Schubeler et al. 2004).
In contrast, another study using ChIP–chip demonstrated in S. cerevisiae that, although H3K79me3 is enriched within transcribed gene regions, there was no correlation between the level of H3K79me3 and transcriptional activity (Pokholok et al. 2005). This was further supported by Zhao and colleagues (Barski et al. 2007) using ChIP coupled with deep sequencing (ChIP-seq) to map mono-, di-, and trimethyl H3K79 in human CD4+ T cells. They found a slight correlation between H3K79me1 and gene activation. However, H3K79me3 was more closely associated with gene repression, while H3K79me2 did not correlate with transcriptional status (Barski et al. 2007).
These two genome-wide analyses were inconsistent with previous studies linking H3K79 methylation to active gene transcription (Ng et al. 2003a; Schubeler et al. 2004; Okada et al. 2005, 2006; Vakoc et al. 2006).

Check 2017 ChIP Validated Histone antibodies
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 H3k79 methylation and other histone modifications.

  • H2B-K123 ubiquitination and H3K79 methylation: deletion of Rad6 and members of the Paf1 complex not only blocked H2B ubiquitination, but also prevented H3K4 and H3K79 methylation
  • In rat, H3K79 methylation also correlates with H4 hyperacetylation but not with active RNA polymerase II, which might point towards a conserved function in chromatin remodeling during the histone-to-protamine transition in both Drosophila and rat.

 

Check other 2017 validated Histone H3 product guide:


4. Related protein study: Dot1 catalyzes the methylation of histone H3 at lysine 79 (H3K9). (1)

Dot1 (also known as KMT4) is an evolutionarily conserved enzyme that catalyzes the methylation of histone H3 at lysine 79. It was initially discovered as a gene that disrupts telomeric silencing when overexpressed (2). Dot1 is the sole enzyme responsible for all forms of H3K79 methylation (mono-methylation, di-methylation, and tri-methylation) in Saccharomyces cerevisiae, Drosophila melanogaster, and humans, based on the fact that knockout of its gene in these organisms results in complete loss of H3K79 methylation (3). The protein is conserved in mammals, called Dot1-like protein (DOT1L), and displays enzymatic properties similar to that of its yeast homologue. Dot1 is unique in being the only non-SET domain containing methyltransferase identified to date (3-5). Two features of this enzyme distinguish it from other known HMTs. First, Dot1 requires a chromatin substrate, and is not active on free histones. Second, the lysine residue modification occurs within the globular region of histone H3, away from the usual histone modification sites that are located in the N- and C-terminal domains. H3K79 is located in the globular domain of histone H3, but it is exposed on the nucleosome surface where it can be methylated by DOT1L. However, this methylation occurs only in a nucleosomal context that is suggestive of a cross talk between H3K79 methylation and other histone modifications (5,6). Studies regarding activity and regulation of Dot1 in many organisms have revealed roles in different biological processes such as transcription, cell-cycle regulation, and DNA damage response.

Check DOT1L antibody

  • Anti-DOT1L Antibody

    $99.00$319.00

    Reactivity: Human
    Applications: ELISA,WB,IHC
    Conjugation: Various
    Supplier: CUSABIO BIOTECH CO.

Source:

  • Farooq Z, Banday S, Pandita TK, Altaf M. The many faces of histone H3K79 methylation. Mutation research Reviews in mutation research. 2016;768:46-52. doi:10.1016/j.mrrev.2016.03.005.
  • Nguyen AT, Zhang Y. The diverse functions of Dot1 and H3K79 methylation. Genes & Development. 2011;25(13):1345-1358. doi:10.1101/gad.2057811.

Reference

  1. Nguyen, A.T., and Zhang, Y., (2011) The diverse functions of Dot1 and H3K79 methylation. Genes & Dev. 2011. 25: 1345-1358
  2. Singer MS, Kahana A, Wolf AJ, Meisinger LL, Peterson SE, Goggin C, Mahowald M, Gottschling DE. Identification of high-copy disruptors of telomeric silencing in Saccharomyces cerevisiae. Genetics. 1998;150:613–632.
  3. VanLeeuwen F, Gafken PR, Gottschling DE. Dot1p modulates silencing in yeast by methylation of the nucleosome core. Cell. 2002;109:745–756.
  4. Ng HH, Feng Q, Wang H, Erdjument-Bromage H, Tempst P, Zhang Y, Struhl K. Lysine methylation within the globular domain of histone H3 by Dot1 is important for telomeric silencing and Sir Protein association. Genes Dev. 2002;16:1518–1527.
  5. Lacoste N, Utley RT, Hunter JM, Poirier GG, Côte J. Disruptor of telomeric silencing-1 is a chromatin-specific histone H3 methyltransferase. J. Biol. Chem. 2002 Aug;277(34):30421–30424.
  6. Feng Q, Wang H, Ng HH, Erdjument-Bromage H, Tempst P, Struhl K, Zhang Y. Methylation of H3-lysine 79 is mediated by a new family of HMTases without a SET domain. Curr. Biol. 2002;12:1052–1058.