Mitochondrial and nucleocytoplasmic isoforms of O-linked GlcNAc transferase encoded by a single mammalian gene
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Mouse tissue distribution of OGT
The Mouse Multiple Choice Northern blots were purchased from OriGene Technologies (Rockville, MD). The blots were prehybridized in 1% bovine serum albumin, 0.5 M NaPO4, pH 7.0, 1 mM EDTA, 7% sodium dodecyl sulfate, and 100 μg/ml denatured salmon testis DNA at 55 °C for 2 h and then hybridized overnight at 55 °C with the gel-purified [α-32P]ATP-radiolabeled human ogt EcoRV–XbaI fragment and the mouse OGT PCR probe (350 bp). The blots were washed twice for 15 min with 0.5% bovine serum albumin, 5% sodium
Tissue distribution of mouse OGT
To examine the relative size and abundance of ogt transcripts in various mouse tissues, Northern blot analysis was performed (Fig. 1). The human ogt probe identified several distinct bands between 9.5 and 4 kb which are present in different amounts in the various mouse tissues tested. Heart and muscle exhibited a relative enrichment of the 6.5 kb species. The exception was stomach, which contained very little of the ogt transcripts. A mouse ogt PCR probe corresponding to the 5′ end of a mouse
Mammalian ogt is highly conserved and is predicted to contain two promoters
The genes encoding the human and rodent OGT are highly conserved in both overall structure and sequence. Several promoter prediction algorithms we employed suggest that ogt may contain two promoters. The most upstream of these contains a prototypic CpG island motif and likely represents the “housekeeping” promoter for OGT. The sequence 1000 bp upstream of the presumptive transcription start contains a large number of transcription factor binding sites. The putative downstream promoter would be
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