Leader peptide efficiency correlates with signal recognition particle dependence in Saccharomyces cerevisiae.
Secretion of bovine pancreatic trypsin inhibitor (BPTI) in Saccharomyces cerevisiae was examined with four different leader peptides: the invertase signal peptide, the mfalpha1 signal peptide, a synthetic signal peptide, and a synthetic pre pro leader. BPTI secretion from a low-copy CEN plasmid varies from 1.8 to 10.4 microgram/mL among these constructs. Secretion titers correlate with dependence on signal recognition particle (SRP), with greatest secretion from the most SRP-dependent construct. Examination of co- vs post-translational translocation pathways and overall translocation efficiency by ubiquitin translocation assay (UTA) does not provide insight into the variation in BPTI secretion efficiency, perhaps due to alteration in translocation kinetics from the additional polypeptide fusion required by the assay. BPTI translocation efficiency (as measured by UTA) is found to drop markedly upon depletion of Srp54p, prior to any observable growth defect. Subsequent to stress response induction and the onset of slow growth (15-h doubling time), BPTI translocation efficiency recovers to the level observed prior to SRP depletion.
Arnold
C E
CE
Department of Chemical Engineering, University of Illinois, Urbana, Illinois 61801, USA.
Parekh
R N
RN
Yang
W
W
Wittrup
K D
KD
D000595:Amino Acid Sequence; D000818:Animals; D007611:Aprotinin / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular / Q000379:methods; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D010957:Plasmids; D021382:Protein Sorting Signals / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000235:genetics; D018271:Signal Recognition Particle / Q000378:metabolism*; D014452:Ubiquitins / Q000378:metabolism
D018840:HSP70 Heat-Shock Proteins; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D018271:Signal Recognition Particle; D014452:Ubiquitins; D007611:Aprotinin