Difference between revisions of "Part:BBa K1051279"

 
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<partinfo>BBa_K1051279 short</partinfo>
 
<partinfo>BBa_K1051279 short</partinfo>
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<h3>Purpose</h3>
 
The measurement of D-box.
 
The measurement of D-box.
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<br />
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<h3>Principle</h3>
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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).
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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.<br />
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                              http://mol-biol4masters.masters.grkraj.org/html/Cell_Cycle_And_Its_Regulation_files/image022.jpg
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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.
  
 
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<span class='h3bb'>Sequence and Features</span>
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<h3>Sequence and Features</h3>
 
<partinfo>BBa_K1051279 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1051279 SequenceAndFeatures</partinfo>
  
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<partinfo>BBa_K1051279 parameters</partinfo>
 
<partinfo>BBa_K1051279 parameters</partinfo>
 
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<h3>References</h3>
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[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;<br />
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[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.<br />

Revision as of 22:44, 27 September 2013

The measurement of D-box.

Purpose

The measurement of D-box.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 150
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1246


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.