title: CD43 gene expression is mediated by a nuclear factor which binds pyrimidine-rich single-stranded DNA.
abstract: CD43 is a leukocyte-specific surface molecule which plays an important role both in adhesion and signal transduction. We have identified a site spanning nucleotides +18 to +39 within the human CD43 gene promoter which in vitro is hypersensitive to cleavage by nuclease S1. Repeats of this region are sufficient to activate expression of a heterologous promoter in CD43-positive cell lines. Two nuclear factors, PyRo1 and PyRo2, interact with the hypersensitive site. PyRo1 is a single-stranded DNA-binding protein which binds the pyrimidine-rich sense strand. Mutation analysis demonstrates that the motif TCCCCT is critical for PyRo1 interaction. Replacement of this motif with the sequence CATATA abolishes PyRo1 binding and reduces expression of the CD43 promoter by 35% in Jurkat T lymphocytic cells and by 52% in the pre-erythroid/pre-megakaryocytic cell line K562. However, this same replacement failed to affect expression in U937 monocytic cells or in CEM T lymphocytic cells. PyRo1, therefore, exhibits cell-specific differences in its functional activity. Further analysis demonstrated that PyRo1 not only interacts with the CD43 gene promoter but also motifs present within the promoters of the CD11a, CD11b, CD11c and CD11d genes. These genes encode the alpha subunits of the beta2 integrin family of leukocyte adhesion receptors. Deletion of the PyRo1 binding site within the CD11c gene reduced promoter activity in T lymphocytic cells by 47%. However, consistent with our analysis of the CD43 gene, the effect of this same deletion within U937 monocytic cells was less severe. That PyRo1 binds preferentially to single-stranded DNA and sequences within the CD43 and CD11 gene promoters suggests that expression of these genes is influenced by DNA secondary structure.
authors.affiliation: Leukocyte Biology and Inflammation Program, Renal Unit, Massachusetts General Hospital, Harvard Medical School, Boston, USA.
authors.forename: O C
authors.initials: OC
authors.lastname: Farokhzad
authors.forename: J M
authors.initials: JM
authors.lastname: Teodoridis
authors.forename: H
authors.initials: H
authors.lastname: Park
authors.forename: M A
authors.initials: MA
authors.lastname: Arnaout
authors.forename: C S
authors.initials: CS
authors.lastname: Shelley
mesh_terms: D015703:Antigens, CD; D001665:Binding Sites; D018845:CD11 Antigens / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000235:genetics; D006801:Humans; D007962:Leukocytes / Q000378:metabolism*; D051917:Leukosialin; D009687:Nuclear Proteins / Q000032:analysis / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011743:Pyrimidines / Q000032:analysis; D012795:Sialoglycoproteins / Q000235:genetics / Q000378:metabolism*; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000378:metabolism
chemical_list: D015703:Antigens, CD; D018845:CD11 Antigens; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D051917:Leukosialin; D009687:Nuclear Proteins; D009838:Oligodeoxyribonucleotides; D011743:Pyrimidines; C465014:SPN protein, human; D012795:Sialoglycoproteins; D015719:Single-Strand Specific DNA and RNA Endonucleases