Regulatory

Part:BBa_K203125

Designed by: Nao Iwamoto and Tim Heinemann   Group: iGEM09_Heidelberg   (2009-10-19)
Revision as of 18:56, 21 October 2009 by Larsplus (Talk | contribs)

SREBP regulated promoter, predicted, tested HB8

SREBP-upregulated promoter predicted by [http://2009.igem.org/Team:Heidelberg/HEARTBEAT HEARTBEAT] (see also [[Part:BBa_K203125:Part_Design|Part Design). SREBP induction experimentally confirmed.


Usage and Biology

SREBP induction. Upon Sterol depletion, the interaction of SCAP and Insig in the ER membrane is inhibited, resulting in cleavage of SREBP, and migration to the nucleus. [http://en.wikipedia.org/wiki/File:WikF1.png Image is public domain.]

Sterol regulatory element-binding protein (SREBP) is a transcription factor involved in the regulation of sterol metabolism. In cells with high concentration of cholestrol SREBP is present in an inactive form anchored to the endoplasmatic reticulum or the nuclear envelop. If the cholesterol concentration decreases SREBP is cleaved by the proteases site-1 protease and site-2 protease resulting in a release of the aminoterminal domain of SREBP. Two additional proteins (Scap and Insig) are needed to regulate this process in a way that the cleavage occurs exclusively during lack of sterol. The aminoterminal domain of SREBP is translocated into the nucleus and binds to the DNA consensus sequence TCACNCCAC. The binding causes an upregulation of the genes needed for cholesterol synthesis. For references, see [http://2009.igem.org/Team:Heidelberg/Eukaryopedia#SREBP Eukaryopedia].

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]

Functional Parameters

Underwent rough characterization by automated plate fluorescence reader/TECAN. Shown to be upregulated by Sterol depletion approx. 8-fold, as compared to cells in a surplus of Sterol and LDL. Corresponds to clone HB8.

HB8 (this part) is upregulated by sterol depletion ("active" condition) approx. 8-fold, as compared to a surplus of sterol. Measured by TECAN / automated plate fluorescence reader. Background was subtracted. For details on induction, refer to [http://2009.igem.org/Team:Heidelberg/Notebook_MaM#Induction Material and Methods]
[edit]
Categories
Parameters
None