Abstract
To understand more about the human inter-alpha-trypsin inhibitor heavy chain H3 (ITIH3) expression and the relationship between this gene and the family of other ITI heavy chain genes, an analysis of the structure of the ITIH3 gene and its promoter region was performed. This gene is a single copy gene, 14 kilobase pair in length and consists of 22 exons. ITIH3 shares highly conserved exon size and intron-exon borders with other ITI heavy chain genes. We determined that the human ITIH1, ITIH3, and ITIH4 genes are closely linked within a 45-kilobase pair. They are arranged in the order of H1-H3-H4, with the ITIH4 gene transcribed in the opposite direction. A model for the evolution of the ITI heavy chain gene family is presented that involves multiple rounds of gene duplication plus inversion events. The minimum promoter region (-135 to +75) is identified in HepG2 cells. The transient transfection study in various cell lines indicates that the activity of the ITIH3 promoter is not liver-specific. DNase I footprinting, mobility shift assays, and cotransfection experiments reveal a functional CCAAT/enhancer-binding protein site (C/EBP, -1344 to -1305) which interacts with C/EBPalpha and C/EBPbeta factors. The latter factors control the transcription of the ITIH3 gene positively.
Highlights
To understand more about the human inter-␣-trypsin inhibitor heavy chain H3 (ITIH3) expression and the relationship between this gene and the family of other ITI heavy chain genes, an analysis of the structure of the ITIH3 gene and its promoter region was performed
Cloning and Sequencing of the ITIH3 Gene—During the initial screening of a human genomic library in EMBL-3 for the ITI heavy chain gene, 10 positive phage clones were identified with heavy chain cDNAs
They showed that the insert of clone G3 contained the 3Ј region of the ITIH1 and the 5Ј region of ITIH3 but lacked the central and 3Ј region of the ITIH3 gene, whereas two clones G21 and G22 were overlapping and contained the 3Ј region coding sequence of ITIH3 (Fig. 1), plus additional genomic DNA extending 7 kb downstream from the exonic boundaries of the gene
Summary
To understand more about the human inter-␣-trypsin inhibitor heavy chain H3 (ITIH3) expression and the relationship between this gene and the family of other ITI heavy chain genes, an analysis of the structure of the ITIH3 gene and its promoter region was performed. To understand the mechanisms regulating the ITIH3 expression and the relationship between the ITIH3 gene and the family of other ITI heavy chain genes, we undertook a structural and functional analysis of the human ITIH3 gene and its promoter region.
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