Expression of FSH in CHO cells, I. Comparison of promoter types and effects of their respective inducers.
The expression levels of recombinant human follicle stimulating hormone (r-hFSH) were studied in several CHO-Kl derived cell lines. The cell lines varied in the promoter used to control r-hFSH expression and in the subunit gene copy ratio. FSH is a heterodimeric molecule, with 2 N-glycosylation sites per peptide chain, and shares a common alpha subunit with the other gonadotropins. Serum stimulated FSH production in the beta actin promoter cell lines 2-3 times over the 7-10 ng/10(6) cells/h levels obtained in protein-free medium. Serum seemed to have roles other than purely at the transcriptional level judging by the increased free alpha to dimer ratio secreted from cells cultured in serum-free medium. Zinc induced FSH expression in metallothionein cell lines, with a 3-fold induction at 50 microM concentrations compared to 0 microM zinc, giving specific productivities of about 7-10 ng/10(6) cells/h, but the induction kinetics were complicated, and suggested other roles for zinc in addition to activation of the metallothionein promoter. Evidence suggested significant post-transcriptional regulation of FSH expression.
Gebert
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Department of Biotechnology, University of NSW, Kensington, Australia.
Gray
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PP
D000199:Actins / Q000235:genetics; D000818:Animals; D001769:Blood; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D005640:Follicle Stimulating Hormone / Q000096:biosynthesis* / Q000235:genetics; D005786:Gene Expression Regulation; D006801:Humans; D008668:Metallothionein / Q000235:genetics; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis; D015032:Zinc / Q000494:pharmacology
D000199:Actins; D011994:Recombinant Proteins; D005640:Follicle Stimulating Hormone; D008668:Metallothionein; D015032:Zinc