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Structure and expression of the mRNA for murine granulocyte‐macrophage colony stimulating factor.
Author(s) -
Gough N.M.,
Metcalf D.,
Gough J.,
Grail D.,
Dunn A.R.
Publication year - 1985
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1002/j.1460-2075.1985.tb03678.x
Subject(s) - holy grail , complementary dna , biology , cdna library , colony stimulating factor , microbiology and biotechnology , gene , genetics , haematopoiesis , computer science , stem cell , world wide web
A cDNA containing a virtually complete copy of the mRNA for the haemopoietic growth regulator, granulocyte‐macrophage colony stimulating factor (GM‐CSF), has been isolated from a murine T lymphocyte cDNA library. When a eukaryotic expression vector with this cDNA coupled to the SV40 late promoter was introduced into simian COS cells, significant quantities of GM‐CSF were secreted. Since all of the biological activities previously ascribed to highly purified GM‐CSF were exhibited in the COS cell‐derived GM‐CSF, all of these activities are intrinsic to the product of a single gene. There are two potential translational initiation codons in the GM‐CSF mRNA; the first is buried in the stem and the second located in the loop of a very stable hairpin structure. Expression studies using deletion derivatives of the cDNA indicated that the second AUG is able to initiate the translation and secretion of GM‐CSF. The amino acid sequence of the leader peptide is rather atypical for a secreted protein and we speculate that molecules which initiate at the first AUG might exist as integral membrane proteins whereas those initiating at the second are secreted.