Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

RNF40 (D2R2O) Rabbit mAb #12187

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP H Endogenous 130 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

RNF40 (D2R2O) Rabbit mAb recognizes endogenous levels of total RNF40 protein. This antibody does not cross-react with RNF20/BRE1A.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asp537 of human RNF40 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using RNF40 (D2R2O) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing Myc/DDK-tagged full-length human RNF20 (hRNF20-Myc/DDK; +) and Myc/DDK-tagged full-length human RNF40 (hRNF40-Myc/DDK; +), using RNF40 (D2R2O) Rabbit mAb (upper) or DYKDDDDK Tag Antibody #2368 (lower).

IP

IP

Immunoprecipitation of RNF40 from 293T cell extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or RNF40 (D2R2O) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using RNF40 (D2R2O) Rabbit mAb.


Background

In mammalian cells, the significance of histone H2B ubiquitination in chromatin epigenetics came from the identification of the budding yeast protein Bre1 (1,2). Together with the ubiquitin-conjugating enzyme Rad6, Bre1 serves as the E3 ligase in the monoubiquitination of the yeast histone H2B within transcribed regions of chromatin (1-3). Subsequently, the mammalian orthologs of yeast Bre1, RNF20 and RNF40, were identified (4,5). These two proteins form a tight heterodimer that acts as the major E3 ligase responsible for histone H2B monoubiquitination at Lys120 in mammalian cells, a modification linked to RNA Pol II-dependent transcription elongation in undamaged cells. Researchers have shown that DNA double-strand breaks (DSBs) are also capable of inducing monoubiquitination of H2B. This process depends upon the recruitment to DSB sites, as well as ATM-dependent phosphorylation of the RNF20-RNF40 heterodimer, thus highlighting a role for this E3 ligase in DSB repair pathways (6). Indeed, investigators have shown that loss of RNF20-RNF40 function promotes replication stress and chromosomal instability, which may constitute an early step in malignant transformation that precedes cell invasion (7).

  1. Wood, A. et al. (2003) Mol Cell 11, 267-74.
  2. Hwang, W.W. et al. (2003) Mol Cell 11, 261-6.
  3. Kao, C.F. et al. (2004) Genes Dev 18, 184-95.
  4. Kim, J. et al. (2005) Mol Cell 20, 759-70.
  5. Zhu, B. et al. (2005) Mol Cell 20, 601-11.
  6. Moyal, L. et al. (2011) Mol Cell 41, 529-42.
  7. Chernikova, S.B. et al. (2012) Cancer Res, Epub ahead of print.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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