The nucleotide sequence of rabbit liver transferrin cDNA (original) (raw)
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Cloning, structural organization and tissue-speci¢c expression of the rabbit transferrin gene
We cloned the rabbit transferrin (rTf) cDNA and gene, and quantified the expression of the rTf gene at the RNA level in various organs. The tissue-specific pattern of expression of rTf gene is different to those in other species, with a high expression in mammary gland and kidney. The exon/intron structure of the rTf gene (17 exons/16 introns) is similar to those of transferrins from other species. The sequence of the rTf cDNA already published is corrected and lengthened in the 5P region, and a likely polymorphism is documented.
Cloning, structural organization and tissue-specific expression of the rabbit transferrin gene
Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression, 1998
We cloned the rabbit transferrin (rTf) cDNA and gene, and quantified the expression of the rTf gene at the RNA level in various organs. The tissue-specific pattern of expression of rTf gene is different to those in other species, with a high expression in mammary gland and kidney. The exon/intron structure of the rTf gene (17 exons/16 introns) is similar to those of transferrins from other species. The sequence of the rTf cDNA already published is corrected and lengthened in the 5P region, and a likely polymorphism is documented.
Cloning and functional expression of the rabbit transferrin gene promoter
The transferrin gene is expressed in all mammals, mainly in the liver. A rabbit genomic library was screened using cDNA probes, and 8 kb of 5∞ flanking sequence of the rabbit transferrin gene was cloned upstream of the cat reporter gene. The first 200 nucleotides of this promoter were sequenced. The rabbit transferrin promoter is highly homologous to the human and murine ones. Its functional activity was tested in the human hepatic cell line HepG2. Using transitory transfections in these cells, a proximal positive region, a negative region and a distal positive region located between −3.6 and −4.0 kb were identified. This distal positive region sequence is highly conserved with the the human gene enhancer sequence, and contains an HNF3a binding site, the mutation of which totally abolished its effect in HepG2 and HuH7 cell lines. The rabbit transferrin 5∞ flanking sequence thus shows a promoter organization very similar to that of the human gene, and the HNF3a binding site in the distal positive region presents the same functional importance in both genes.
The Journal of biological chemistry, 1991
We present a comparative study of the cis- and trans-acting elements governing the expression of the human transferrin (Tf) gene in two tissues, liver and testis, where Tf is expressed at various levels. We have previously identified the elements of the promoter, negative, and enhancer regions involved in the liver-specific expression of the gene. By transfection experiments of primary cultured rat Sertoli cells compared with hepatoma cells, DNase I footprinting, and gel retardation studies, we have analyzed 3.6 kilobase pairs of the Tf regulatory region. The far upstream enhancer functional in Hep3B cells is inactive in Sertoli cells; in the two cell types, different nuclear factors appear to bind to a DNA domain crucial for enhancer activity. Similar negative- and positive-acting elements are present in the distal promoter in both tissues. However different combinations of proximal promoter elements control tissue-specific expression. Liver-specific transcription is governed by th...
The complete amino acid sequence of human serum transferrin
Proceedings of the National Academy of Sciences, 1982
The complete amino acid sequence of human serum transferrin has been determined by aligning the structures of the 10 CNBr fragments. The order of these fragments in the polypeptide chain is deduced from the structures of peptides overlapping methionine residues and other evidence. Human transferrin contains 678 amino acid residues and--including the two asparagine-linked glycans--has an overall molecular weight of 79,550. The polypeptide chain contains two homologous domains consisting of residues 1-336 and 337-678, in which 40% of the residues are identical when aligned by inserting gaps at appropriate positions. Disulfide bond arrangements indicate that there are seven residues between the last half-cystine in the first domain and the first half-cystine in the second domain and therefore, a maximum of seven residues in the region of polypeptide between the two domains. Transferrin--which contains two Fe-binding sites--has clearly evolved by the contiguous duplication of the structural gene for an ancestral protein that had a single Fe-binding site and contained approximately 340 amino acid residues. The two domains show some interesting differences including the presence of both N-linked glycan moieties in the COOH-terminal domain at positions 413 and 610 and the presence of more disulfide bonds in the COOH-terminal domain (11 compared to 8). The locations of residues that may function in Fe-binding are discussed.
Synthesis and Secretion of Transferrin by Cultured Mouse Hepatoma Cells
Differentiation, 1978
The mouse hepatoma cell (Hepa-1) in tissue culture has been shown to synthesize and secrete three electrophoretically distinct transfemns. Each of these forms of transferrin has a molecular weight of 77,000, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The concentration of each form is indicated by its staining intensity, which is highest in the form with the fastest mobility and lowest in the form with the slowest mobility. The relative rate of transferrin synthesis has been determined in log-phase and stationary-phase cells; the data indicate that the relative rate of synthesis increases twofold in stationary-phase cells. When the incorporation of [3Hlleucine into transferrin reaches steady state, the rate of secretion is equal to the rate of synthesis; the rate of secretion also increases twofold in stationary-phase cells. Our studies also show that transfemn synthesis accounts for 0.98% of the total protein synthesis in log-phase cells and for 1.8% in stationary-phase cells. This is the level of synthesis that has been determined by in vivo studies. We conclude that after continuous culture for several years these hepatoma cells have maintained one of the characteristics of the differentiated liver cell, namely, the ability to synthesize and secrete transferrin.
Biochemistry, 1990
A human liver cDNA library was screened with a synthetic oligonucleotide, complementary to the 5' region of human transferrin mRNA, as a hybridization probe. The full-length human cDNA clone isolated from this screen contained part of the 5' untranslated region, the complete coding region for the signal peptide and the two lobes of transferrin, the 3' untranslated region, and a poly(A) tail. By use of oligonucleotide-directed mutagenesis in vitro, two translational stop codons and a Hind111 site were introduced after the codon for Asp-337. This fragment was inserted into two different expression vectors that were then introduced into Escherichia coli. As judged by NaDodSO4-po1yacrylamide gel electrophoresis and Western blot analysis, however, recombinant hTF/2N was undetectable in bacteria transformed by these plasmids. Concurrently, we developed a plasmid vector for the expression of recombinant hTF/2N in eukaryotic cells. In this case, a DNA fragment coding for the natural signal sequence, the hTF/2N lobe, I Abbreviations: hTF, human serum transferrin; hTF/2N, aminoterminal half-molecule of hTF; hGH, human growth hormone; BHK, cultured baby hamster kidney cells; DHFR, dihydrofolate reductase; DMEM, Dulbecco's modified essential medium; FPLC, fast protein liquid chromatography; MTX, methotrexate; NMR, nuclear magnetic resonance; NTA, nitrilotriacetate; PAGE, polyacrylamide gel electrophoresis.
Evidcnce for the Functional Heterogeneity of the Two Sites of Transferrin in Vitro
British Journal of Haematology, 1980
ABSTRACT A recently developed crossed immunoelectrophoretic method for displaying and quantitating the four possible molecular species of transferrin has been utilized to assess the relative effectiveness of each site of rabbit and human diferric transferrin in providing iron to rabbit reticulocytes. The site which appears to reside in the N-terminal half of the rabbit protein was found to be at least 5 times more effective than its counterpart. However, both sites may serve as iron donors in monoferric as well as diferric rabbit transferrins. It is also possible that iron may be removed from rabbit transferrin in pairwise as well as sequential fashion. In human diferric transferrin, the site in the C-terminal domain functions as the better iron donor for rabbit reticulocytes.