SignalSilence® PLK1 siRNA I #6292
Supporting Data
| REACTIVITY | H |
Species Cross-Reactivity Key:
- H-Human
Product Information
Product Usage Information
CST recommends transfection with 100 nM SignalSilence® PLK1 siRNA I 48 to 72 hours prior to cell lysis. For transfection procedure, follow protocol provided by the transfection reagent manufacturer. Please feel free to contact CST with any questions on use.
Each vial contains the equivalent of 100 transfections, which corresponds to a final siRNA concentration of 100 nM per transfection in a 24-well plate with a total volume of 300 μl per well.
Storage
Product Description
Quality Control
Oligonucleotide synthesis is monitored base by base through trityl analysis to ensure appropriate coupling efficiency. The oligo is subsequently purified by affinity-solid phase extraction. The annealed RNA duplex is further analyzed by mass spectrometry to verify the exact composition of the duplex. Each lot is compared to the previous lot by mass spectrometry to ensure maximum lot-to-lot consistency.
Background
At least four 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). In addition, 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; DNA damage prevents phosphorylation at these sites (14).
- Nigg, E.A. (1998) Curr Opin Cell Biol 10, 776-83.
- Toyoshima-Morimoto, F. et al. (2002) EMBO Rep 3, 341-8.
- Toyoshima-Morimoto, F. et al. (2001) Nature 410, 215-20.
- Peter, M. et al. (2002) EMBO Rep 3, 551-6.
- Jackman, M. et al. (2003) Nat Cell Biol 5, 143-8.
- Nakajima, H. et al. (2003) J Biol Chem 278, 25277-80.
- Sumara, I. et al. (2002) Mol Cell 9, 515-25.
- Hauf, S. et al. (2001) Science 293, 1320-3.
- Peters, J.M. (1999) Exp. Cell Res. 248, 339-49.
- Kraft, C. et al. (2003) EMBO J 22, 6598-609.
- Kotani, S. et al. (1998) Mol Cell 1, 371-80.
- Jang, Y.J. et al. (2002) J Biol Chem 277, 44115-20.
- Smits, V.A. et al. (2000) Nat Cell Biol 2, 672-6.
- Tsvetkov, L. and Stern, D.F. (2005) Cell Cycle 4, 166-71.
限制使用
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