The beta7 integrin gene (Itgb-7) promoter is responsive to TGF-beta1: defining control regions.
The beta7 integrins LPAM-1 (alpha4beta7) and M290 (alphaEbeta7) mediate the homing of lymphocytes to gut-associated lymphoid tissue, and the proposed retention of intraepithelial lymphocytes (IEL), respectively. Here we show that the gut mucosal cytokine TGF- beta1 increases the expression of beta7 and alphaE subunit mRNA transcripts and the cell-surface expression of M290 on T cells, and that it decreases the level of alpha4 integrin transcripts. Induced beta7 integrin gene expression was inhibited by the protein tyrosine kinase inhibitor genistein, implicating a role for tyrosine phosphorylation. An analysis of the beta7 integrin gene promoter revealed three DNAse I hypersensitivity sites, two of which mapped to the 5' and 3' ends of a promoter fragment (nucleotides +690 to +63) that directed both the basal and the TGF-beta1-induced expression of a heterologous reporter gene. Deletion analysis identified two TGF-beta1 response regions encompassing nucleotides -509 to -398 (TGFBRR1), and -122 to +32 (TGFBRR2). TGFBRR1 interacted with at least five protein complexes, whose binding could be induced with TGF-beta1 stimulation and could be antagonized by TGFBRR2 which harbored both similar and distinctive cis-elements. TGFBRR2 interacted specifically with at least two major nuclear protein complexes, whose binding was phosphorylation dependent. These data provide new insights into the mechanism by which TGF-beta may switch LPAM-1(+ve) migrating T cells to express M290, facilitating their retention in the gut.
Department of Molecular Medicine, School of Medicine and Health Science, University of Auckland, Auckland, New Zealand.
S P
SP
Lim
E
E
Leung
G W
GW
Krissansen
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D004247:DNA / Q000235:genetics; D003850:Deoxyribonuclease I; D039641:Integrin beta Chains; D016023:Integrins / Q000737:chemistry / Q000235:genetics*; D007413:Intestinal Mucosa / Q000166:cytology / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism; D015345:Oligonucleotide Probes / Q000235:genetics; D010766:Phosphorylation; D011401:Promoter Regions, Genetic / Q000187:drug effects*; D011487:Protein Conformation; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D014443:Tyrosine / Q000235:genetics / Q000378:metabolism
D039641:Integrin beta Chains; D016023:Integrins; D009687:Nuclear Proteins; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; C071777:integrin alpha4beta7; C070709:integrin alphaEbeta7; C091368:integrin beta7; D014443:Tyrosine; D004247:DNA; D003850:Deoxyribonuclease I