Part:BBa_K203106

HMG-CoA synthase promoter (sterol regulated)

This part is a natural HMG CoA synthase promoter responsive to SREBP and when placed upstream of other genes could induce their transcription in response to lipid addition to the medium.

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 up regulation of the genes needed for cholesterol synthesis. See reference at [http://2009.igem.org/Team:Heidelberg/Eucaryopedia#Transcription_factors Eucaryopedia].

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

n/a	HMG-CoA synthase promoter (sterol regulated)

The natural HMG CoA synthase promoter was induced by Lipoprotein Deficient Serum (more details: see Cell Culture Notebook) in HeLa cells. Promoter activtiy was then roughly characterized analogous to REU by TECAN (automated plate fluorescence reader). For more natural promoters look at Part:BBa_K203105 and Part:BBa_K203107.

BBa_K203106 is a natural HMG CoA synthase promoter induced approximately 35% by Lipoprotein Deficient Serum. The GFP flourescence of the HMG CoA synthase promoter was measured by TECAN once (à three replicates). Background fluorescence was substracted, and fluorescence levels are plotted relative to Part:BBa_K203112.