Activation of the gamma-glutamyltransferase promoter 2 in the rat colon carcinoma cell line CC531 by histone deacetylase inhibitors is mediated through the Sp1 binding motif.
The single-copy gene for rat gamma-glutamyltransferase (GGT) encodes at least seven distinct mRNAs that differ in their 5'-untranslated regions only. Tissue- and developmental-specific expression of GGT is partly achieved by the presence of many transcription factor-binding sites in the promoters of this gene. In an earlier study we found that GGT mRNAs II and IV levels were increased upon butyrate-induced differentiation of the rat colon carcinoma cell line CC531. The mechanism for this butyrate-induced upregulation remains unknown, but may result from altered promoter activity as butyrate is a known histone deacetylase inhibitor. In the present study, we show by transient transfection studies that butyrate enhanced the expression of the luciferase reporter gene driven by the rat GGT promoter 2 (P2). Trichostatine A (TSA), another histone deacetylase inhibitor, also enhanced transcription from this promoter. The role of the transcription factor site Sp1 in butyrate- or TSA-induced activation of the GGT P2 was examined as Sp1 has been previously shown to play a central role in the transcriptional activation of other genes during butyrate and TSA stimulation. A triple sequence-motif of this isolated Sp1 site linked to a minimal promoter was able to mediate butyrate- and TSA-induced expression of the luciferase reporter gene, while no effect was measured using the minimal promoter alone. Deleting the Sp1 site in the context of the rat GGT P2 strongly reduced the basal transcription activity and abrogated butyrate- and TSA-induced activation of the mutated promoter. These results suggest that butyrate- or TSA-induced activation of the rat GGT P2 can be mediated by a Sp1 binding motif.
Mikkelsen
Idun Merete
IM
Department of Medical Biochemistry, Institute of Medical Biology, Faculty of Medicine, University of Tromsø, N-9037 Tromsø, Norway. idunm@fagmed.uit.no
Huseby
Nils-Erik
NE
Visvikis
Athanase
A
Moens
Ugo
U
D000818:Animals; D001483:Base Sequence; D002087:Butyrates / Q000494:pharmacology*; D003110:Colonic Neoplasms / Q000473:pathology; D004247:DNA / Q000032:analysis; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009154:Mutation; D011401:Promoter Regions, Genetic / Q000187:drug effects* / Q000235:genetics; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D016329:Sp1 Transcription Factor / Q000378:metabolism; D014407:Tumor Cells, Cultured; D005723:gamma-Glutamyltransferase / Q000235:genetics / Q000378:metabolism*
D002087:Butyrates; D004791:Enzyme Inhibitors; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D012333:RNA, Messenger; D016329:Sp1 Transcription Factor; C012589:trichostatin A; D004247:DNA; D005723:gamma-Glutamyltransferase