@entity6585 is expressed in many different cell types, and serves a variety of regulatory functions. In order to better understand the transcriptional control of this protein, the 5' promoter region of the @entity1 @entity6585 gene was cloned and sequenced. After confirming the transcription start point (TSP) using 5' rapid amplification of cDNA ends analysis, the 5' promoter was analysed. The sequence lacks a TATA box, but contains a variety of putative regulatory elements. There are two GAS sites, two @entity1178 sites, two overlapping Sp-1 sites, and a gamma-IRE site clustered between -1080 and -1450. There is another cluster of putative regulatory sites between the TSP and -550 which contains two Sp-1 sites, two @entity18720 sites, one GAS site, one NF-kappaB site, an incomplete CAAT box (8/9) and an overlapping Sp-1/ @entity18720 site at -17 to -26. Reporter gene constructs containing 4225 and 1498 bases 5' of the TSP demonstrated excellent unidirectional transcriptional activity in both constructs. Reporter genes containing serial 5' deletions were compared to the -1498 construct. The reporter gene which contained base pairs (bp) -36 to +89 had almost no activity. The reporter gene containing -188 to +89 had 50% of the -1498 construct, indicating that this sequence contains at least the minimal promoter. The Sp-1/ @entity18720 site near the transcription start site was studied by electrophoretic mobility shift and reporter gene assays. Addition of HeLa cell nuclear extract to labeled double-stranded (ds) oligonucleotide containing this sequence resulted in a gel shift which was inhibited by excess unlabeled ds oligonucleotide and by a consensus cold Sp-1 ds oligonucleotide, indicating specific Sp-1 binding. Excess @entity18720 or NF-kappaB ds oligonucleotide had no effect on nuclear protein binding to the sequence. Mutation of the @entity6585 wild-type Sp-1/ @entity18720 sequence eliminated both nuclear protein binding and the sequences ability to compete with native sequence for nuclear binding protein. A -1048 to +89 reporter construct containing a mutated Sp-1/ @entity18720 site resulted in a 40% decrease in transcriptional activity. Therefore, the 5' flanking sequence of the @entity6585 gene exhibits promoter activity which may be localized to a variety of controlling regions, of which the proximal Sp-1/ @entity18720 site appears to be important for basal activity via its Sp-1 binding ability.

Characterization of the XXXX @entity6585 promoter sequence.

@entity1 :: ('9606', 'Species') :: ['human'] @entity6585 :: ('6281', 'Gene') :: ['p11'] @entity1178 :: ('3725', 'Gene') :: ['AP-1'] @entity18720 :: ('7020', 'Gene') :: ['AP-2']
@entity1 :: (9606,Species) :: ['human']
