Cloning and nucleotide sequence of DNA coding for bovine preproparathyroid hormone.

Kronenberg, Henry M.; McDevitt, Barbara E.; Majzoub, Joseph A.; Nathans, Jeremy; Sharp, Phillip A.; Potts, John T.; Rich, Alexander

doi:10.1073/pnas.76.10.4981

Cited by 96 publications

(29 citation statements)

References 29 publications

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“…1, right). These fragments were compatible with the published DNA sequence (10,11) and suggested that our clone included the entire coding region for preproPTH. (10) showing cell-free translation products of poly (A)-rich RNA in a wheat germ system (6) (Fig.…”

Section: Resultssupporting

confidence: 60%

Direct regulation by calcium of cytoplasmic messenger ribonucleic acid coding for pre-proparathyroid hormone in isolated bovine parathyroid cells.

Russell¹,

Lettieri²,

Sherwood³

1983

J. Clin. Invest.

141

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A B S T R A C T DNA complementary to bovine preproparathyroid hormone mRNA was cloned and labeled by nick translation in order to measure mRNA by molecular hybridization. Bovine parathyroid cells were maintained in primary tissue culture for periods up to 96 h at 0.5 mM, 1.25 mM, and 2.5 mM calcium, which was followed by extraction of cellular RNA. Levels of mRNA showed no differences at 0.5 or 1.25 mM calcium, but at high calcium levels, there was a reversible decrease that began at 16 h to a plateau at 30% of control after 72 h. These studies suggest that the glandular capacity to synthesize hormone may be at or near maximal at normal calcium, but at high calcium, there is a decrease over time in steady state levels of mRNA.

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Section: Resultssupporting

confidence: 60%

Direct regulation by calcium of cytoplasmic messenger ribonucleic acid coding for pre-proparathyroid hormone in isolated bovine parathyroid cells.

Russell¹,

Lettieri²,

Sherwood³

1983

J. Clin. Invest.

141

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“…This is consistent with the hypothesis that the preference of cytidine over uridine in the third position of codons minimizes translation errors caused by codon-anticodon base wobble (33). The codon preferences in bovine prothrombin mRNA are generally similar to those preferred in other bovine mRNAs (34,35). DNA sequence analysis of the flanking region shows that the prothrombin cDNA is inserted in the same orientation as the ampicillin gene of the pBR322 cloning vector.…”

Section: A G a G G C C A A T C C A G T G A A T G A A T T A T T T T supporting

confidence: 71%

Cloning and analysis of a cDNA coding for bovine prothrombin.

MacGillivray¹,

Degen²,

Chandra³

et al. 1980

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

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Poly(A)RNA enriched for prothrombin was isolated by specific immunoprecipitation ofbvine liver polysomes. Prothrombin consisted of about 8% of the cell-free translation products of this RNA. A double-stranded cDNA was synthesized by using reverse transcriptase (RNA-dependent DNA nucleotidyltransferase) and made blunt-ended with nuclease SI. After tailing with dCTP and terminal transferase, the double-stranded cDNA was annealed to pBR322 DNA that had been cleaved previously at the single Pst I site and similarly tailed with dGTP. The resulting plasmids were used to transform Escherichia coli strain RRI under P3-EKI conditions. Sixtythree tetracycline-resistant clones were obtained that hybridized to nP-labeled cDNA synthesized from prothrombin-enriched mRNA. Recombinants containing cDNA to prothrombin mRNA sequences were screened by a solution hybridization assay with a[cDNA synthesized from mRNA. This enriched mRNA was 50% prothrombin mRNA, as determined by a reticulocyte lysate translation assay. Three positive clones were identified by this assay; they contained bovine DNA inserts of 700,500, and 400 base pairs. The DNA sequence of the 700-base-pair insert was then determined. This recombinant plasmid contained DNA coding for the carboxyl-terminal 160 residues of bovine prothrombin followed by a noncoding region of 119 base pairs and a poly(A) tail of 60 base pairs.The coagulation of blood is the result of a series of stepwise reactions in which many of the coagulation proteins in plasma are converted from zymogens to highly specific serine proteases (see ref. 1 for a recent review). In the final stages of the blood coagulation cascade, prothrombin is converted to thrombin by Factor Xa in the presence of phospholipid, calcium ions, and Factor Va. Thrombin then cleaves fibrinogen to form an insoluble fibrin clot. Although prothrombin is one of the most abundant coagulation proteins present in plasma, its mRNA constitutes only about 1% of the total poly(A)-RNA in bovine liver (2). This level of RNA makes it difficult to purify prothrombin mRNA to the degree of homogeneity required for many biochemical investigations. Recent developments in recombinant DNA techniques, however, permit the isolation of large amounts of homogeneous DNA corresponding to the specific mRNAs. Such cloned DNAs can be used for many different studies, such as the quantitation of cellular mRNA levels in response to various stimuli, the purification of specific mRNA by hybridization to immobilized DNA, and as a hybridization probe to isolate and study genomic DNA.In this paper, we describe the construction, cloning in Escherichia colh, and sequence of a recombinant plasmid containing DNA coding for the carboxyl-terminal portion of bovine prothrombin. Enrichment of mRNA. Polysomes were prepared from bovine liver as described (2). Polysomes synthesizing prothrombin were enriched by specific immunoprecipitation with antiprothrombin prepared in rabbits and either goat anti-rabbit immunoglobulin coupled to p-aminobenzyl-cellulose (3) or ...

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“…Restriction (4). Double-stranded cDNA with poly(dC) homopolymeric extensions was synthesized as described (9)(10)(11)(12).…”

mentioning

confidence: 99%

Nucleotide sequence of the mRNA encoding the pre-alpha-subunit of mouse thyrotropin.

Chin¹,

Kronenberg²,

Dee³

et al. 1981

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

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We have constructed and cloned in bacteria recombinant DNA molecules containing DNA sequences coding for the precursor of the a subunit of thyrotropin (pre-TSH-a). Double-stranded DNA complementary to total poly(A)+RNA derived from a mouse pituitary thyrotropic tumor was prepared enzymatically, inserted into the Pst I site of the plasmid pBR322 by using poly(dC)-poly(dG) homopolymeric extensions, and cloned in Escherichia coli x1776. Cloned cDNAs encoding pre-TSH-a were identified by their hybridization to pre-TSH-a mRNA as determined by cell-free translations of hybrid-selected and hybrid-arrested RNA. The nucleotide sequences oftwo cDNAs (510 and 480 base pairs) were determined with chemical methods and corresponded to much of the region coding for the a subunit and the 3' untranslated region of pre-TSH-a mRNA. The sequence of the 5' end of the mRNA was determined from cDNA synthesized by using total mRNA as template and a restriction enzyme DNA fragment as primer. Together these sequences represented >90% of the coding and noncoding regions of full-length pre-TSH-a mRNA, which was determined to be 800 bases long. The amino acid sequence of the pre-TSH-a deduced from the nucleotide sequence showed a NH2-terminal leader sequence of24 amino acids followed by the 96-amino-acid sequence ofthe apoprotein of TSHa. There is greater than 90% homology in the amino acid sequences among the murine, ruminant, and porcine a subunits and 75-80% homology among the murine, equine, and human a subunits. Several regions of the sequence remain absolutely conserved among all species, suggesting that these particular regions are essential for the biological function of the subunit. The successful cloning of the a subunit of TSH will permit further studies of the organization of the genes coding for the glycoprotein hormone subunits and the regulation of their expression.

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Cloning and nucleotide sequence of DNA coding for bovine preproparathyroid hormone.

Cited by 96 publications

References 29 publications

Direct regulation by calcium of cytoplasmic messenger ribonucleic acid coding for pre-proparathyroid hormone in isolated bovine parathyroid cells.

Direct regulation by calcium of cytoplasmic messenger ribonucleic acid coding for pre-proparathyroid hormone in isolated bovine parathyroid cells.

Cloning and analysis of a cDNA coding for bovine prothrombin.

Nucleotide sequence of the mRNA encoding the pre-alpha-subunit of mouse thyrotropin.

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