Part:BBa_K1503000

Cell cycle dependend degradation-tag from S. cerevisia.

This degradation-tag, in short D-tag, is a short part of the cyclin Clb3. It is the sequence for the amino acids of the N-terminal until and including a destruction box, a nine amino acid recognition site with the sequence R-x-x-L-x-x-x-x-N. This sequence is necessary but not sufficient for a specific ubiqitin-mediated degradation and to be found in most of the B-cyclins of yeast [1].
Cyclins bind to cell cycle specific kinases and mediate thereby the progress of the different steps of the cell cycle. The six B-cyclins in yeast (Clb1-6) are responsible for the transition into S- and M-Phase of the cell cycle [2]. With increasing digit the degradation occours earlier in the cell cycle [3]. As we reviewed the sequences of the cyclins it became obvious that the distance of the destruction box to the N-terminal of the protein increases with the digit, stating an inverse relation [4].

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 94
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

UV picture of the gel with the amplified d-tag fragment from S. cerevisiae.

References

1. Zachariae W., Destruction with a box: substrate recognition by the anaphase-promoting complex. Mol Cell. 2004 Jan 16;13(1):2-3.

2. Fitch I, et al. (1992) Characterization of four B-type cyclin genes of the budding yeast Saccharomyces cerevisiae. Mol Biol Cell 3(7):805-18.

3. Arellano M, Moreno S., Regulation of CDK/cyclin complexes during the cell cycle. Int J Biochem Cell Biol. 1997 Apr;29(4):559-73.

4. Sequences of the Clb proteins to be found in the Yeast Genome Database