Abstract
Signal sequences are evolutionarily conserved and are often functionally interchangeable between prokaryotes and eukaryotes. However, we have found that the bacterial signal peptide, OmpA, functions incompletely in insect cells. Upon baculovirus-mediated expression of chloramphenicol acetyltransferase (CAT) in insect cells, OmpA signal peptide led to the cytosolic accumulation of the CAT molecules in an aglycosylated, signal-peptide cleaved form, in addition to the secretion of the glycosylated CAT. When green fluorescent protein (GFP) was used as another reporter, the GFP molecules expressed from the OmpA-GFP construct was distributed primarily in the cytosol as aggresome-like structures. These results together suggest that, subsequent to the cleavage of OmpA signal peptide in the ER, some of the processed proteins are returned to the cytoplasm. Since the prototypical insect signal peptide, melittin, did not result in this ER-to-cytosol dislocation of the reporter proteins, we proposed a model explaining the dislocation process in insect cells, apparently selective to the OmpA-directed secretory pathway bypassing the co-translational transport.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 131-142 |
| Number of pages | 12 |
| Journal | Cell Structure and Function |
| Volume | 28 |
| Issue number | 2 |
| State | Published - Apr 2003 |
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Keywords
- Aggresome
- Baculovirus
- ER dislocation
- Post-translational transport
- Signal peptide
ASJC Scopus subject areas
- Physiology
- Structural Biology
- Cell Biology
Cite this
Functional characterization of bacterial signal peptide OmpA in a baculovirus-mediated expression system. / Nagamine, Toshihiro; Kawasaki, Yu; Iizuka, Tetsutaro; Okano, Keiju; Matsumoto, Shogo; Choudary, Prabhakara V.
In: Cell Structure and Function, Vol. 28, No. 2, 04.2003, p. 131-142.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Functional characterization of bacterial signal peptide OmpA in a baculovirus-mediated expression system
AU - Nagamine, Toshihiro
AU - Kawasaki, Yu
AU - Iizuka, Tetsutaro
AU - Okano, Keiju
AU - Matsumoto, Shogo
AU - Choudary, Prabhakara V
PY - 2003/4
Y1 - 2003/4
N2 - Signal sequences are evolutionarily conserved and are often functionally interchangeable between prokaryotes and eukaryotes. However, we have found that the bacterial signal peptide, OmpA, functions incompletely in insect cells. Upon baculovirus-mediated expression of chloramphenicol acetyltransferase (CAT) in insect cells, OmpA signal peptide led to the cytosolic accumulation of the CAT molecules in an aglycosylated, signal-peptide cleaved form, in addition to the secretion of the glycosylated CAT. When green fluorescent protein (GFP) was used as another reporter, the GFP molecules expressed from the OmpA-GFP construct was distributed primarily in the cytosol as aggresome-like structures. These results together suggest that, subsequent to the cleavage of OmpA signal peptide in the ER, some of the processed proteins are returned to the cytoplasm. Since the prototypical insect signal peptide, melittin, did not result in this ER-to-cytosol dislocation of the reporter proteins, we proposed a model explaining the dislocation process in insect cells, apparently selective to the OmpA-directed secretory pathway bypassing the co-translational transport.
AB - Signal sequences are evolutionarily conserved and are often functionally interchangeable between prokaryotes and eukaryotes. However, we have found that the bacterial signal peptide, OmpA, functions incompletely in insect cells. Upon baculovirus-mediated expression of chloramphenicol acetyltransferase (CAT) in insect cells, OmpA signal peptide led to the cytosolic accumulation of the CAT molecules in an aglycosylated, signal-peptide cleaved form, in addition to the secretion of the glycosylated CAT. When green fluorescent protein (GFP) was used as another reporter, the GFP molecules expressed from the OmpA-GFP construct was distributed primarily in the cytosol as aggresome-like structures. These results together suggest that, subsequent to the cleavage of OmpA signal peptide in the ER, some of the processed proteins are returned to the cytoplasm. Since the prototypical insect signal peptide, melittin, did not result in this ER-to-cytosol dislocation of the reporter proteins, we proposed a model explaining the dislocation process in insect cells, apparently selective to the OmpA-directed secretory pathway bypassing the co-translational transport.
KW - Aggresome
KW - Baculovirus
KW - ER dislocation
KW - Post-translational transport
KW - Signal peptide
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UR - http://www.scopus.com/inward/citedby.url?scp=0038420737&partnerID=8YFLogxK
M3 - Article
VL - 28
SP - 131
EP - 142
JO - Cell Structure and Function
JF - Cell Structure and Function
SN - 0386-7196
IS - 2
ER -