| Autor: |
Hjelmsoe, Iben, Allen, Carl E., Cohn, Martin A., Tulchinsky, Eugene M., Wu, Lai-Chu |
| Zdroj: |
Journal of Biological Chemistry; January 2000, Vol. 275 Issue: 2 p913-920, 8p |
| Abstrakt: |
A κB-like sequence, Sb, is integral to the composite enhancer located in the first intron of the metastasis-associated gene, S100A4/mts1. Oligonucleotides containing this sequence form three specific complexes with nuclear proteins prepared from S100A4/mts1-expressing CSML100 adenocarcinoma cells. Protein studies show the Sb-interacting complexes include NF-κB/Rel proteins, p50·p50 and p50·p65 dimers. Additionally, the Sb sequence was bound by an unrelated ∼200-kDa protein, p200. Site-directed mutagenesis in conjunction with transient transfections indicate that p200, but not the NF-κB/Rel proteins, transactivates S100A4/mts1. To identify candidate genes for p200, double-stranded DNA probes containing multiple copies of Sb were used to screen a randomly primed λgt11 cDNA expression library made from CSML100 poly(A)+RNA. Two clones corresponding to the DNA-binding proteins KRCand Alf1were identified. KRCencodes a large zinc finger protein that binds to the κB motif and to the signal sequences of V(D)J recombination. In vitroDNA binding assays using bacterially expressed KRC fusion proteins, demonstrate specific binding of KRC to the Sb sequence. In addition, introduction of KRCexpression vectors into mammalian cells induces expression ofS100A4/mts1and reporter genes driven byS100A4/mts1gene regulatory sequences. These data indicate that KRCpositively regulates transcription ofS100A4/mts1. |
| Databáze: |
Supplemental Index |
| Externí odkaz: |
|
