#Q16763
27338
Product Information
Product Usage Information
CST recommends transfection with 100 nM SignalSilence® UBE2S siRNA II 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
Specificity / Sensitivity
Species Reactivity:
Human
Product Description
Background
Protein ubiquitination requires the concerted action of the E1, E2, and E3 ubiquitin-conjugating enzymes. Ubiquitin is first activated through ATP-dependent formation of a thiol ester with ubiquitin-activating enzyme E1. The activated ubiquitin is then transferred to a thiol group of ubiquitin-carrier enzyme E2. The final step is the transfer of ubiquitin from E2 to an ε-amino group of the target protein lysine residue, which is mediated by ubiquitin-ligase enzyme E3 (1).
The human anaphase promoting complex (APC/C) is a large macromolecular E3 ligase complex that is largely responsible for the timely progression through mitosis via the sequential targeting of cell cycle regulators for proteasomal degradation. Recent work has revealed that APC/C substrates are marked for proteasomal degradation during cell cycle progression through the covalent assembly of Lys11-linked ubiquitin chains, which occurs through a priming phase and elongation phase (2-5). The APC/C utilizes, in part, the UBE2C/UBCH10 E2 enzyme to prime substrates for degradation through the covalent attachment of short Lys11-linked chains (3,6). The Lys11-specific elongating E2 enzyme, UBE2S/E2-EPF, extends these short chains into long Lys11-linked ubiquitin chains on APC/C bound substrates (2,3,7).
In addition to the well-established biochemical role for UBE2S in cell cycle regulation, this enzyme has been shown to be overexpressed in many types of human cancer (8) and has also been implicated in hypoxia signaling (9,10). Indeed, UBE2S has been reported to associate with VHL and to target it for proteasomal degradation, thereby stabilizing HIF-1α (9).
- Hershko, A. (1988) J Biol Chem 263, 15237-40.
- Williamson, A. et al. (2009) Proc Natl Acad Sci USA 106, 18213-8.
- Jin, L. et al. (2008) Cell 133, 653-65.
- Wu, T. et al. (2010) Proc Natl Acad Sci USA 107, 1355-60.
- Song, L. and Rape, M. (2010) Mol Cell 38, 369-82.
- Summers, M.K. et al. (2008) Mol Cell 31, 544-56.
- Wickliffe, K.E. et al. (2011) Cell 144, 769-81.
- Tedesco, D. et al. (2007) Neoplasia 9, 601-13.
- Jung, C.R. et al. (2006) Nat Med 12, 809-16.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
Limited Uses
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