Effect of elongation factor 1α promoter and SUMF1 over in vitro expression of N-acetylgalactosamine-6-sulfate sulfatase

• DOI: 10.1007/s11033-008-9392-3
• Publication Date: 2008-11-06T16:22:14Z
• Authors: Carlos J. Almécig...; Maria A. Rueda-Pa...; Angela J. Espejo; Olga Y. Echeverri; Adriana Montaño; Shunji Tomatsu; Luis A. Barrera
• Keywords: 0301 basic medicine; Binding Sites; Sp1 Transcription Factor; Computational Biology; Cytomegalovirus; Gene Expression; Transfection; Chondroitinsulfatases; Cell Line; 03 medical and health sciences; Peptide Elongation Factor 1; Humans; CpG Islands; Oxidoreductases Acting on Sulfur Group Donors; Sulfatases; Promoter Regions, Genetic; Plasmids

Morquio A is an autosomal recessive disease caused by the deficiency of N-acetylgalactosamine-6-sulfate sulfatase (GALNS), leading to the lysosomal accumulation of keratan-sulfate and chondroitin-6-sulfate. We evaluated in HEK293 cells the effect of the cytomegalovirus immediate early enhancer/promoter (CMV) or the elongation factor 1alpha (EF1alpha) promoters, and the coexpression with the sulfatase modifying factor 1 (SUMF1) on GALNS activity. Four days postransfection GALNS activity in transfected cells with CMV-pIRES-GALNS reached a plateau, whereas in cells transfected with EF1alpha-pIRES-GALNS continued to increase until day 8. Co-transfection with pCXN-SUMF1 showed an increment up to 2.6-fold in GALNS activity. Finally, computational analysis of transcription factor binding-sites and CpG islands showed that EF1alpha promoter has long CpG islands and high-density binding-sites for Sp1 compared to CMV. These results show the advantage of the SUMF1 coexpression on GALNS activity and indicate a considerable effect on the expression stability using EF1alpha promoter compared to CMV.