Part:BBa_K1051201

D-BOX

Purpose

Works for the degradation of fluorescent proteins we used at N-terminal nearby.

Principle

The best studied substrates of ubiquitin- and APC/C-mediated proteolysis are the mitotic cyclins (Murray, Solomon, & Kirschner, 1989) Mitosis-exit inducer, a cyclin B, are highly dependent on its 90 residues for ubiquitin-mediated proteolysis. Analysis of its N-terminal region demonstrated the sequence essential for cyclin proteolysis, called ‘destruction box’(D-box). Meanwhile, when cyclin destruction started, substrates containing D-boxes were rapidly poly-ubiquitinated. The consensus motif in B-type cyclins is RXALGXIXN. For much of the cell cycle, the D-box may not be recognized with high affinity, and when it is recognized, the ‘bait’ construct becomes highly unstable(Yamano, Tsurumi, Gannon, & Hunt, 1998).

When the D-box exists in the protein N-terminal, this protein is recognized more easily by the E1 and then captured into the ubiquitin-mediated degradation pathway. Thus, we combined it to the fluorescent proteins’ N-terminal to fasten their degradation and avoid veiling the following fluorescence.

                              http://mol-biol4masters.masters.grkraj.org/html/Cell_Cycle_And_Its_Regulation_files/image022.jpg

The said sequences of cyclins are targets for ubiquitination by E1, E2, and E3 by Anaphase Promoting Complex (APC/C) at the end of anaphase. However cyclins-C, F, G and H have structural relationship but not involved in cell cycle regulation

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

References

[1] Zachariae W, Nasmyth K (August 1999). "Whose end is destruction: cell division and the anaphase-promoting complex". Genes Dev. 13 (16): 2039–58. doi:10.1101/gad.13.1;
[2] Yamano H, Tsurumi C, Gannon J, Hunt T. (1998 Oct )The role of the destruction box and its neighbouring lysine residues in cyclin B for anaphase ubiquitin-dependent proteolysis in fission yeast: defining the D-box receptor.6.2039. PMID 10465783.