Expression of a Mutated Bovine Growth Hormone Gene Suppresses Growth of Transgenic Mice

To determine the importance of the third α-helix in bovine growth hormone (bGH) relative to growth-related biological activities, the following experimental approach was used: (i) mutagenesis of helix III of bGH to generate an idealized amphiphilic helix; (ii) in vitro expression analyses of the mut...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1990-07, Vol.87 (13), p.5061-5065
Main Authors: Chen, Wen Y., Wight, David C., Wagner, Thomas E., Kopchick, John J.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Amino acids
Aminoacids, peptides. Hormones. Neuropeptides
Analytical, structural and metabolic biochemistry
Animals
Base Sequence
Biological and medical sciences
Cattle
Cell Membrane
Cultured cells
Female
Fundamental and applied biological sciences. Psychology
Gene Expression
Genes
Growth Hormone - genetics
Growth Hormone - physiology
Kinetics
L cells
L Cells (Cell Line) - metabolism
Liver
Liver - metabolism
Male
Mice
Mice, Transgenic - growth & development
Molecular Sequence Data
Molecules
Mutation
Plasmids
Protein Conformation
Proteins
Receptors
Receptors, Somatotropin - metabolism
Transgenic animals
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Summary:To determine the importance of the third α-helix in bovine growth hormone (bGH) relative to growth-related biological activities, the following experimental approach was used: (i) mutagenesis of helix III of bGH to generate an idealized amphiphilic helix; (ii) in vitro expression analyses of the mutated bGH gene in cultured mouse L cells; (iii) mouse liver membrane binding studies of wild-type and mutated bGH; and (iv) expression of the mutated gene in the transgenic mouse. An altered bGH gene (pBGH10Δ 6-M8) was generated that encodes the following changes: glutamate-117 to leucine, glycine-119 to arginine, and alanine-122 to aspartate. The plasmid pBGH10Δ 6-M8 was shown to be expressed in, and its protein product secreted by, mouse L cells. The altered hormone possessed the same binding affinity to mouse liver membrane preparations as wild-type bGH. Transgenic mice containing the mutated bGH gene, however, showed a significant growth-suppressed phenotype. The degree of suppression was directly related to serum levels of the altered bGH molecule.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.87.13.5061