Down-Regulator of Transcription 1, TBP-Binding Negative Cofactor 2, NC2b

• Species; Human
• Expression Host: E.coli
• Tag: His-tag
• Purity: 90%
• Molecular Weight: 20.6 kDa.
• Gene Accession Number: NM_001938.

Purification and Quality Control

The His-tag recombinant protein is purified by affinity chromatography in combination with FPLC columns. The purified Dr1 is greater than 90% homogeneous based on SDS-PAGE analysis.

Unit Definition (Activity)

1 unit equals 1 nanogram of purified protein. 1 unit is sufficient for a gel mobility shift assay in a 20 µl reaction; 100 units are sufficient for protein-protein interaction assays.

Applications

Dr1 has been applied in reconstituted in vitro transcription assays and protein-protein interactions assays. For Research Use Only.

Formulation and Storage

The protein is in 20mM Tris-HCl pH7.9,100mM NaCl, 0.2mM EDTA, 1mM DTT and 20% glycerol. Stored at -70°C before use. Avoid repeated freeze thaw cycles.

Synonym

down-regulator of transcription 1, TBP-binding negative cofactor 2, NC2, NC2-BETA, NC2b.

Protein Sequence

MASSSGNDDD LTIPRAAINK MIKETLPNVR VANDARELVV NCCTEFIHLI SSEANEICNK SEKKTISPEH VIQALESLGF GSYISEVKEV LQECKTVALK RRKASSRLEN LGIPEEELLR QQQELFAKAR QQQAELAQQE WLQMQQAAQQ AQLAAASASA SNQAGSSQDE EDDDDI

Background

Dr1 is a general negative regulator of class II and class III gene expression (1, 2). It binds to the basic repeat domain of TBP on promoter DNA and can prevent the RNA polymerase II holoenzyme, or its TFIIB and/or TFIIA subunits, from assembling into an initiation complex (3, 4). Dr1 is phosphorylated in vivo and this modification affects its interaction with TBP (1). In addition, Dr1 interacts with the hyperphosphorylated form of Pol II and with the repression domain of the AREB6 repressor (5, 6). Dr1 forms a heterodimer complex with DRAP1 through its histone fold domain (7). More recently it was discovered that Dr1-DRAP1 is a bi-functional basal transcription factor that differentially regulates gene transcription through DPE (downstream promoter elements) or TATA box motifs (8). It can stimulate transcription in vitro from Drosophila promoters containing DPEs, whereas it represses transcription from TATA-containing promoters (8).

References

1. Inostroza, J.A., et al., (1992) Cell 70, 477-489
2. White, R.J., et al., (1994) Science 266, 448-451
3. Mermelstein, F., et al., (1996) Genes Dev. 10, 1033-1048
4. Kim, T.K., et al., (1995) J. Biol. Chem. 270, 10976-10981
5. Castano, E., et al., (2000) Proc. Natl. Acad. Sci. USA 97, 7184-7189
6. Ikeda, K., et al., (1998) Mol. Cell. Biol. 18, 10-18
7. Goppelt, A., et al., (1996) EMBO J. 15, 3105-3116
8. Willy, P. J., et al., (2000) Science 290, 982-9858
9. Denko et al., (2003) J. of Bio. Chem., 278:5744-5749

DISCLAIMER

This products is recommended For RESEARCH USE ONLY and is Not qualified for Use in Diagnostic or Therapeutic Procedures.