title: Induction of 6-sulfated glycans with cell adhesion activity via T-bet and GATA-3 in human helper T cells.
abstract: BACKGROUND
  Cell surface 6-sulfated glycans play important roles in various immunological events through cell-to-cell interactions. The 6-sulfation process is mediated by 6-sulfotransferase family isoenzymes. We previously demonstrated that GlcNAc6ST-1, one of the isoenzyme genes, is induced by GATA-3 and NF-κB in human helper T (Th) cells. However, transcriptional regulation of HEC-GlcNAc6ST, another isoenzyme important in Th cells, remains unclear.
  
  
  METHODS
  5'-RACE analysis, chromatin immunoprecipitation, and reporter assays were performed to reveal transcriptional regulation of HEC-GlcNAc6ST. RNA-knockdown and forced expression experiments were performed to demonstrate the contribution of HEC-GlcNAc6ST to the 6-sulfated glycan expression.
  
  
  RESULTS
  We identified potential binding sites of Sp1, T-bet, and GATA-3 in the HEC-GlcNAc6ST promoter. Reporter assays indicated that transfection of Sp1 enhanced the activity, whereas mithramycin A, an Sp1-specific inhibitor, repressed it. Transfection of T-bet increased the activity, which was inhibited by introducing a mutation into the potential T-bet binding site. GATA-3 alone could not elevate the activity, although co-transfection of protein kinase A, which is known to enhance IL-5 transcription in Th2 cells through phosphorylation of GATA-3, caused elevation. RNA-knockdown and forced expression of HEC-GlcNAc6ST in Jurkat cells down- and up-regulated α2,6-sialylated 6-sulfo N-acetyllactosamine, a preferential ligand for B-cell-specific CD22 antigen, respectively. From these results, we concluded that T-bet and GATA-3 as well as Sp1 control the expression of glycan with cell-adhesion activity by regulating HEC-GlcNAc6ST transcription in Th cells.
  
  
  CONCLUSIONS
  These results may provide a clue to biological regulation of Th-cell interaction with selectins and other carbohydrate-recognition molecules by T-bet and GATA-3.
authors.affiliation: Division of Molecular Pathology, Aichi Cancer Center, Nagoya, Aichi 464-8681, Japan.
authors.forename: Keiichiro
authors.initials: K
authors.lastname: Sakuma
authors.forename: Guo-Yun
authors.initials: GY
authors.lastname: Chen
authors.forename: Masahiro
authors.initials: M
authors.lastname: Aoki
authors.forename: Reiji
authors.initials: R
authors.lastname: Kannagi
mesh_terms: D015153:Blotting, Western; D002448:Cell Adhesion / Q000502:physiology*; D002450:Cell Communication; D047369:Chromatin Immunoprecipitation; D005434:Flow Cytometry; D050990:GATA3 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation; D006801:Humans; D015848:Interleukin-5 / Q000235:genetics / Q000378:metabolism; D019169:Jurkat Cells; D016256:Lewis X Antigen / Q000031:analogs & derivatives; D016328:NF-kappa B / Q000235:genetics* / Q000378:metabolism; D009844:Oligosaccharides / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000080506:Sialyl Lewis X Antigen / Q000031:analogs & derivatives; D015238:Sulfotransferases / Q000235:genetics* / Q000378:metabolism; D020825:T-Box Domain Proteins / Q000235:genetics / Q000378:metabolism*; D006377:T-Lymphocytes, Helper-Inducer / Q000378:metabolism*; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D000097811:Carbohydrate Sulfotransferases; D000098815:T-bet Transcription Factor
chemical_list: D050990:GATA3 Transcription Factor; D015848:Interleukin-5; D016256:Lewis X Antigen; D016328:NF-kappa B; D009844:Oligosaccharides; D012333:RNA, Messenger; D000080506:Sialyl Lewis X Antigen; D015238:Sulfotransferases; D020825:T-Box Domain Proteins; D000097811:Carbohydrate Sulfotransferases; D000098815:T-bet Transcription Factor; C086949:6'-sulfated sialyl Lewis x; C494749:GATA3 protein, human