[A prosegment of the yeast alpha-factor controls a heterologous protein (human growth factor) in Saccharomyces cerevisiae culture media].
The role of the yeast pheromone prosegment--alpha-factor--in the export of the heterologous protein, the human growth hormone (hGH), in the culture medium of S. cerevisiae has been studied. Using genetic engineering constructions, it has been shown that different N-terminal signal peptides (SP) are not able to provide the hGH export. Transformant cells carrying the plasmid with a hybrid sequence encoding the prosegment-hGH (without SP) accumulate non-glycosylated pro-hGH in the cytosol. Only the combination of SP and the prosegment as the N-terminal fragment of the hGH precursor results in the processing and export of the mature form of the hormone. The origin (or type) of SP is of no significance. The glycosylation inhibitors--2-deoxy-D-glucose and tunicamycin--suppress the export but not the entry of hGH into the periplasm, thus indicating a critical role of the intactness of the prosegment polymannose chains for the efficient export of heterologous protein. A conclusion is drawn that the two preprosegment parts play different roles. The SP constituent of prepro-hGH introduces pro-GH into the general secretory pathway, while the prosegment resulting from the SP cleavage directs hGH from the cell into the culture medium.
Tsiomenko
A B
AB
Tuĭmetova
G P
GP
El'darov
M A
MA
Korolev
S V
SV
Skriabin
K G
KG
Kulaev
I S
IS
D000595:Amino Acid Sequence; D001483:Base Sequence; D001692:Biological Transport; D003470:Culture Media; D003847:Deoxyglucose / Q000494:pharmacology; D006031:Glycosylation; D006133:Growth Substances / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D000072235:Mating Factor; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010455:Peptides / Q000737:chemistry / Q000502:physiology*; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000378:metabolism*; D014415:Tunicamycin / Q000494:pharmacology
D003470:Culture Media; D006133:Growth Substances; D009838:Oligodeoxyribonucleotides; D010455:Peptides; D014415:Tunicamycin; D000072235:Mating Factor; D003847:Deoxyglucose
Prosegment drozhzhevogo alpha-faktora napravliaet geterolgichnyĭ belok (gormon rosta cheloveka) v kul'tural'nuiu sredu Saccharomyces cerevisiae.