Molecular cloning and nucleotide sequence of a transforming gene detected by transfection of chicken B-cell lymphoma DNA

Goubin, Gérard; Goldman, Debra S.; Luce, Judith; Neiman, Paul E.; Cooper, Geoffrey M.

doi:10.1038/302114a0

Cited by 188 publications

(60 citation statements)

References 53 publications

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“…Nucleotides encoding the leader sequence of ovotransferrin are 56% homologous to these described here in human Tf. Although the ChBlym-J transforming gene in chicken lymphoma DNA (6) is homologous with the NH2-terminal region of human Tf, its leader sequence bears little, if any, homology with that of Tf.…”

Section: Resultsmentioning

confidence: 99%

Human transferrin: cDNA characterization and chromosomal localization.

Yang¹,

Lum²,

McGill³

et al. 1984

Proc. Natl. Acad. Sci. U.S.A.

276

134

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Transferrin (Tf) is the major iron binding protein in vertebrate serum. It shares homologous amino acid sequences with four other proteins: lactotransferrin, ovotransferrin, melanoma antigen p97, and HuBlym-l. Antigen p97 and the Tf receptor genes have been mapped on human chromosome 3. The goal of the study described here was to initiate the characterization of the Tf gene by identifying and characterizing its cDNA and mapping its chromosomal location. Recombinant plasmids containing human cDNA encoding Tf have been isolated by screening an adult human liver library with a mixed oligonucleotide probe. Within the 2.3 kilobase pairs of TfcDNA analyzed, there is a probable leader sequence encoded by 57 nucleotides followed by 2037 nucleotides that encode the homologous amino and carboxyl domains. During evolution, three areas of the homologous amino and carboxyl domains have been strongly conserved, possibly reflecting functional constraints associated with iron binding. Chromosomal mapping by in situ hybridization and somatic cell hybrid analysis indicate that the Tfgene is located at q21-25 on human chromosome 3, consistent with linkage of the Tf, Tf receptor, and melanoma p97 loci.Transferrin (Tf) carries ferric iron from the intestine, reticuloendothelial system, and liver parenchymal cells to all proliferating cells in the body. The family of Tf-like proteins represents the product of an intragenic duplication followed by a series of independent gene duplications (1-4). Serum Tf (1), hen ovotransferrin (2, 3), lactotransferrin (4), melanoma antigen p97 (5), and a transforming protein from chicken lymphoma ChBlym-1 (6) share strong amino acid sequence homologies. A transforming protein from Burkitt lymphomas recently described (7) may also belong to the Tf family. There is also significant internal homology in the amino-terminal (NH2) and carboxyl-terminal (COOH) domains of Tf, lactotransferrin, and ovotransferrin (1-4). For example, the NH2 and COOH domains of human Tf reveal 40% identity when the NH2 domain (residues 1-336) and the COOH domain (residues 337-678) are compared (1). In the study described Screening of cDNA Clones. The cDNA library, kindly provided by Stuart H. Orkin (Harvard Medical School, Boston) was constructed from human liver RNA as described (10). The cDNA library was incubated overnight on L agar plates containing 10 jig of tetracycline per ml and was transferred to nitrocellulose filters. The plasmids were amplified with 250 ,tg of chloramphenicol per ml and the filters were prepared for hybridization as described by Grunstein and Hogness (11). The filters were hybridized at 37°C with 32P-labeled oligonucleotide mixed probes as described by Wallace, et al. (12). The hybridization mixture contained 0.9 M NaCl, 0.09 M Tris HCl (pH 7.5), 0.006 M EDTA, 0.5% NaDodSO4, Denhardt's (13) solution (5 x strength), 100 ,ug of denatured Escherichia coli DNA per ml, and 6.4 ng of 5' end-labeled oligonucleotide mixed probes per ml having a specific activity of -7 x 108 cpm/,ug.Preparation...

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Section: Resultsmentioning

confidence: 99%

Human transferrin: cDNA characterization and chromosomal localization.

Yang¹,

Lum²,

McGill³

et al. 1984

Proc. Natl. Acad. Sci. U.S.A.

276

134

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“…c-rnwc is amplified in human lung cancer cell lines (28) and highly expressed in cell lines derived from human lung and ovarian tumors (15) growth in what appears to be a sequential manner. It has been known for some time that c-mnyc acts in concert with a second oncogene, c-blym, in virus-induced bursal lymphomas of chickens (20). The same two oncogenes appear to be implicated in the genesis of Burkitt lymphomas as well (11).…”

Section: Resultsmentioning

confidence: 99%

Regulated transcription of c-Ki-ras and c-myc during compensatory growth of rat liver.

Goyette¹,

Petropoulos²,

Shank³

et al. 1984

Mol. Cell. Biol.

205

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We examined the transcription of six cellular oncogenes during the process of compensatory growth in rat liver after partial hepatectomy. We have previously reported that transcripts of c-rasH are elevated during regenerative growth of the liver. We now report that transcripts of c-rasK and c-myc genes are significantly elevated after partial hepatectomy, whereas transcripts of c-abl and c-src are essentially unchanged and transcripts of c-mos are undetectable in either normal or regenerating rat liver. In liver regeneration after partial hepatectomy or chemical injury, changes in c-myc transcripts occur before DNA synthesis. The elevation of c-myc and c-ras transcripts is sequential in that highest levels of c-myc transcripts were detected 12 to 18 h after partial hepatectomy, whereas the levels of c-rasH and c-rasK were maximal by 36 to 48 h. Transcripts of all three activated oncogenes returned to their basal levels by 96 h.

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“…Hence, decreased T f R density appears to be an early single-cell marker for changes in cell growth rate. This is particularly interesting in light of the observed relationships between the ras and XCH Blym-1 mammalian oncogene products and Tf or TfRs (9).…”

Section: Discussionmentioning

confidence: 99%

Single‐cell analysis of the relationship among transferrin receptors, proliferation, and cell cycle phase in K562 cells

et al. 1985

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Multiparameter single-cell analysis by flow cytometry was used to distinguish between size-related changes in K562 cell transferrin receptor (TfR) expression and changes in membrane receptor density throughout the cell cycle and over time in culture. Lightscatter pulse-width time-of-flight, a direct and readily calibrated measure of cell diameter, was used to calculate receptor density as the average number of receptors per unit cell surface area. Cell surface TfRs were unimodally distributed over the cell population and were present throughout the cell cycle. The number of receptors increased as cells progressed through the cell cycle, but cell cycle phase was also correlated with cell volume. However, when size heterogeneity was factored out by reanalysis of listmode data, there was a clear cell-cycle effect: among cells of the same size, both the number of receptors per cell and the receptor density increased from G1 to S to G2IM. TfEt expression was also followed over time in culture after dilution into fresh medium. A decrease in growth rate after four days was preceded by one to two days by a decrease in both number of TfRs per cell and mean receptor density, indicating that decreased TfR expression represented true "down-regulation" and not just decreased cell size or an increase in the proportion of smaller G1 cells. This type of analysis is generally applicable for resolving the effects of cell size heterogeneity and cell cycle on membrane protein distribution and for other studies of ligand-receptor interaction.

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Molecular cloning and nucleotide sequence of a transforming gene detected by transfection of chicken B-cell lymphoma DNA

Cited by 188 publications

References 53 publications

Human transferrin: cDNA characterization and chromosomal localization.

Human transferrin: cDNA characterization and chromosomal localization.

Regulated transcription of c-Ki-ras and c-myc during compensatory growth of rat liver.

Single‐cell analysis of the relationship among transferrin receptors, proliferation, and cell cycle phase in K562 cells

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