Characterization of Ets-binding sequence of human transglutaminase 3 gene promoter

Experimental Dermatology
Chang Deok KimJeung-Hoon Lee

Abstract

The Ets- and Sp1-transcription factors have been implicated in the epithelial specific expression of transglutaminase 3 gene (TGM3) in vitro. By electrophoretic mobility shift assay (EMSA), the core motif of Ets-binding sequence of TGM3 was determined as ACAGGAAT (-118 to -111 bp from transcriptional start site). However, a sequence extending from -120 to +10 of TGM3 proximal promoter region failed to induce the expression of CAT reporter in transfected normal human epidermal keratinocytes (NHEKs). In contrast, a construct extending from -126 to +10 bp showed the highest expression of CAT gene, indicating the presence of an important element(s) between -126 and -120 bp that affects TGM3 expression in NHEKs. To find the critical sequences in this promoter region, we performed EMSA analysis with competitor oligonucleotides, in which the upstream nucleotides of Ets core motif were mutated. The results showed that the mutation of A at -125 dramatically reduced the binding of Ets to its recognition sequence. Transient transfection assay revealed that the mutation in this nucleotide greatly diminished the expression of CAT reporter gene in NHEKs. Together, these results suggest that the upstream sequence of Ets core motif is critical...Continue Reading

References

Jul 28, 1995·The Journal of Biological Chemistry·S Y KimP M Steinert
May 7, 1998·The International Journal of Biochemistry & Cell Biology·A D SharrocksP R Yates
Mar 14, 2000·Molecular and Cellular Biology·S SinhaE Fuchs
Mar 20, 2002·The Journal of Experimental Medicine·Miklós SárdyNeil Smyth
Sep 5, 2002·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Andrey E KalininPeter M Steinert

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Citations

Jul 15, 2005·Skin Pharmacology and Physiology·M J Choi, H I Maibach
Mar 2, 2005·The Journal of Investigative Dermatology·Richard L EckertEllen A Rorke
Dec 9, 2008·Annales de dermatologie et de vénéréologie·V Doffoel-HantzC Bédane

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