Cell Signaling Technology

Product Pathways - DNA Damage

TLK1 Antibody #4125

Applications Reactivity Sensitivity MW (kDa) Source
W IF-IC H M R Endogenous 86 Rabbit

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

TLK1 Antibody detects endogenous levels of total TLK1. The antibody may cross-react with TLK2.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues adjacent to Ser183 of human TLK1. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa (human), C6 (rat), and RAW 264.7 (mouse) cells, using TLK1 Antibody.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using TLK1 Antibody (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Background

Tousled-like kinases (TLK1 and TLK2) are nuclear serine/threonine kinases named for their homology to the Tousled gene from Arabidopsis thaliana, essential for flower development (1). The kinase activities of the TLKs are cell cycle regulated, with maximal activity during S phase (1). TLK appears to play a role in chromatin assembly and DNA damage checkpoint regulation (1,2). In C. elegans, TLK1 is essential for appropriate transcription during embryonic development (3). Substrates for TLK include the human chromatin assembly factor Asf, which functions in DNA replication- and repair-coupled chromatin assembly (2). DNA damage during S phase, when TLK is maximally active, leads to inhibition of TLK activity (1). This inhibition requires ataxia mutated kinase (ATM) and Chk1 (4,5). ATM and the related kinase ATR are activited by DNA damage during S phase, phosphorylate Chk1/Chk2, and block the transition into mitosis (6). Chk1 phosphorylates TLK1 on Ser695 in vitro and in vivo, leading to inhibition of TLK1 activity (4). This process likely provides a mechanism to slow the chromatin assembly processes controlled by TLK in the event of DNA damage.

  1. Sillje, H. H. et al. (1999) EMBO J. 18, 5691-5702.
  2. Sillje, H.H. and Nigg, E.A. (2001) Curr. Biol. 11, 1068-1073.
  3. Han, Z. et al. (2003) Curr. Biol. 13, 1921-1929.
  4. Groth, A. et al. (2003) EMBO J. 22, 1676-1687.
  5. Krause, D. R. et al. (2003) Oncogene 22, 5927-5937.
  6. Kastan, M.B. and Lim, D.S. (2000) Nat. Rev. Mol. Cell Biol. 1, 179-186.

Application References

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Companion Products

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

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