Part:BBa_K243028

FOS-Split Linker-Fok_a

Activator protein-1 (AP-1) is a crucial transcription factor implicated in numerous cancers and binds DNA as a dimer. Two classes of core DNA sequences, the sequences TRE (TGACTCA) and CRE (TGACGTCA), can be recognized by the AP-1. Nine homologues of the AP-1 leucine zipper region have been characterized. All of them are able to form heterodimers, some also form homodimers. One of them is the so called c-Fos. Via leucin zipper they interact among each other and with their bipartite domain they bind DNA. (Abate et al., Mol Cell Biol. 1991 July).

Heterodimeric Fos complex (http://www.rcsb.org/pdb/explore/images.do?structureId=1FOS)

Usage and Biology

To avoid the labeling of oligos we tried an alternative way of binding of the heterodimeric Fok to the DNA by using the DNA binding domain of Fos as an adapter between the Fok construct and the DNA. For our needs, we chose the natural bZIP sequence of Fos and Jun.

We fused Split Linker-Fok_a to Fos.The mixture of this construct with another hybrid protein called Jun allows the dimerization of Fos and therefore its binding at their target sequence TRE (TGACTCA) or CRE (TGACGTCA) by using the bZIP sequence of Jun as adapter. The addition of Fok_i leads to DNA cleavage.

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 19
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 766