DNA

Part:BBa_K1051201

Designed by: wanling ZHOU   Group: iGEM13_Shenzhen_BGIC_ATCG   (2013-06-30)

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.

                              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.

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Categories
Parameters
n/aD-BOX