Part:BBa_K626008

DBD designed for RARE in ADRB - EcR LBD

Composite construct built from zinc finger polymer designed to bind to RARE in adrenergic beta-1- receptor and Ecdysone receptor Ligand Bining Domain (EcR LBD).

Zinc finger

Zinc fingers are small protein structural motifs binding one or more zinc ions to help stabilize their folds. Zinc finger proteins often bind to DNA and RNA because their shape allows close interaction of the domain with the nucleotides of DNA and RNA, and they can also bind to proteins and small molecules. Their nucleotide binding properties allow them to function in regulating gene expression and in virus assembly.

Zinc fingers coordinate zinc ions with a combination of cysteine and histidine residues. They can be classified by the type and order of these zinc coordinating residues (e.g., Cys2His2, Cys4, and Cys6). A more systematic method classifies them into different "fold groups" based on the overall shape of the protein backbone in the folded domain. The most common "fold groups" of zinc fingers are the Cys2His2-like (the "classic zinc finger"), treble clef, and zinc ribbon. The DNA binding domain of nuclear hormone receptors is a highly conserved domain containing two zinc fingers that binds to specific sequences of DNA called hormone response elements (HRE).

EcR LBD

The ecdysone receptor is a nuclear receptor found in arthropods, where it controls development and contributes to other processes such as reproduction. The receptor is a non-covalent heterodimer of two proteins, the EcR protein and ultraspiracle protein (USP). It binds to and is activated by ecdysteroids. Pulses of 20-hydroxyecdysone occur during insect development, whereupon this hormone binds to the ecdysone receptor, a ligand-activated transcription factor found in the nuclei of insect cells. This in turn leads to the activation of many other genes, which ultimately causes physiological changes that result in ecdysis (moulting).

More on Ecdysone receptor here: [http://en.wikipedia.org/wiki/Ecdysone_receptor].

Ecdysone receptor Uniprot: [http://www.uniprot.org/uniprot/P34021].

View the Ecdysone receptor crystal structure here: [http://www.rcsb.org/pdb/explore/jmol.do?structureId=1R1K&bionumber=1]

The part functions as a nuclear hormone receptor, hence can be applied as a regulator of expression of a gene with a response element to the zinc finger.

pSB1C3-ADRB-ECR-LBD and pSB1C3-CD38-ECR-LBD (circular and NotI digested).

Sequence and Features

Assembly Compatibility:

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

  Illegal BglII site found at 1135
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]