Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

PLK4 Antibody #3258

Applications Reactivity Sensitivity MW (kDa) Source
W IF-IC H M R Mk Endogenous 95 Rabbit

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

Protocols

Specificity / Sensitivity

PLK4 Antibody detects endogenous levels of total PLK4 protein.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Cys458 of human PLK4. Antibodies were purified by peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of recombinant GST-PLK4 #7580 and extracts from MOLT4, RL and Ramos cell lines using PLK4 Antibody.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HT-1080 cells using PLK4 Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Background

At least 4 distinct polo-like kinases exist in mammalian cells: PLK1, PLK2, PLK3, and PLK4/SAK (1). PLK1 apparently plays many roles during mitosis, particularly in regulating mitotic entry and exit. The mitosis promoting factor (MPF), cdc2/cyclin B1, is activated by dephosphorylation of cdc2 (Thr14/Tyr15) by cdc25C. PLK1 phosphorylates cdc25C at Ser198 and cyclin B1 at Ser133 causing translocation of these proteins from the cytoplasm to the nucleus (2-5). PLK1 phosphorylation of Myt1 at Ser426 and Thr495 has been proposed to inactivate Myt1, one of the kinases known to phosphorylate cdc2 at Thr14/Tyr15 (6). Polo-like kinases also phosphorylate the cohesin subunit SCC1, causing cohesin displacement from chromosome arms that allow for proper cohesin localization to centromeres (7). Mitotic exit requires activation of the anaphase promoting complex (APC) (8), a ubiquitin ligase responsible for removal of cohesin at centromeres, and degradation of securin, cyclin A, cyclin B1, Aurora A, and cdc20 (9). PLK1 phosphorylation of the APC subunits Apc1, cdc16, and cdc27 has been demonstrated in vitro and has been proposed as a mechanism by which mitotic exit is regulated (10,11).Substitution of Thr210 with Asp has been reported to elevate PLK1 kinase activity and delay/arrest cells in mitosis, while a Ser137Asp substitution leads to S-phase arrest (12). Additionally, while DNA damage has been found to inhibit PLK1 kinase activity, the Thr210Asp mutant is resistant to this inhibition (13). PLK1 has been reported to be phosphorylated in vivo at Ser137 and Thr210 in mitosis, and DNA damage prevents phosphorylation at these sites (14).

PLK4/SAK is transcriptionally repressed by p53 and may contribute to p53-mediated apoptosis (12). PLK4 has also been identified as a key regulator of centriole duplication (13-15).

  1. Nigg, E.A. (1998) Curr. Opin. Cell Biol. 10, 776-783.
  2. Toyoshima-Morimoto, F. et al. (2002) EMBO Rep. 3, 341-348.
  3. Toyoshima-Morimoto, F. et al. (2001) Nature 410, 215-220.
  4. Peter, M. et al. (2002) EMBO Rep. 3, 551-556.
  5. Jackman, M. et al. (2003) Nat. Cell Biol. 5, 143-148.
  6. Nakajima, H. et al. (2003) J. Biol. Chem. 278, 25277-25280.
  7. Sumara, I. et al. (2002) Mol. Cell 9, 515-525.
  8. Hauf, S. et al. (2001) Science 293, 1320-1323.
  9. Peters, J.M. (1999) Exp. Cell Res. 248, 339-349.
  10. Kraft, C. et al. (2003) EMBO J. 22, 6598-6609.
  11. Kotani, S. et al. (1998) Mol. Cell 1, 371-380.
  12. Jang, Y.J. et al. (2002) J Biol Chem 277, 44115-20.
  13. Smits, V.A. et al. (2000) Nat Cell Biol 2, 672-6.
  14. Tsvetkov, L. and Stern, D.F. (2005) Cell Cycle 4, 166-71.
  15. Li, J. et al. (2005) Neoplasia 7, 312-323.
  16. Habedanck, R. et al. (2005) Nat. Cell Biol. 7, 1140-1146.
  17. Ko, M.A. et al. (2005) Nat. Genet. 37, 883-888.
  18. Bettencourt-Dias, M. et al. (2005) Curr. Biol. 15, 2199-2207.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

Products