scholarly journals XBP1 is Critical to Protect Cells from Endoplasmic Reticulum Stress: Evidence from Site-2 Protease-deficient Chinese Hamster Ovary Cells

2006 ◽  
Vol 31 (2) ◽  
pp. 117-125 ◽  
Author(s):  
Hiderou Yoshida ◽  
Satomi Nadanaka ◽  
Ryuichiro Sato ◽  
Kazutoshi Mori
Keyword(s):  
Endoplasmic Reticulum ◽  
Endoplasmic Reticulum Stress ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Ovary Cells ◽  
Protease Deficient

Reduction of endogenous GRP78 levels improves secretion of a heterologous protein in CHO cells

1988 ◽  
Vol 8 (10) ◽  
pp. 4063-4070
Author(s):  
A J Dorner ◽  
M G Krane ◽  
R J Kaufman
Keyword(s):  
Endoplasmic Reticulum ◽  
Chinese Hamster Ovary ◽  
Heterologous Protein ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Glycosylation Site ◽  
Ovary Cells ◽  
Tissue Plasminogen ◽  
Stable Association

GRP78 is localized in the endoplasmic reticulum and associates with improperly folded or underglycosylated proteins. The role of GRP78 in secretion was studied in Chinese hamster ovary cells expressing a tissue plasminogen activator (tPA) variant which lacks potential N-linked glycosylation site sequences because of mutagenesis. The expression of variant tPA resulted in elevated levels of GRP78 and its stable association with tPA. The introduction of antisense GRP78 genes resulted in a two- to threefold reduction in GRP78 levels compared with those of the original cells. Cells with reduced levels of GRP78 secreted two- to threefold-higher levels of tPA activity. tPA expressed in these cells displayed reduced association with GRP78, and a greater proportion was processed to the mature form and secreted. These results demonstrate that reduction of GRP78 level can improve the secretion of an associated protein.

scholarly journals Analysis of ATF6 Activation in Site-2 Protease-deficient Chinese Hamster Ovary Cells

2006 ◽  
Vol 31 (2) ◽  
pp. 109-116 ◽  
Author(s):  
Satomi Nadanaka ◽  
Hiderou Yoshida ◽  
Ryuichiro Sato ◽  
Kazutoshi Mori
Keyword(s):  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Ovary Cells ◽  
Protease Deficient

scholarly journals DistinctC-mannosylation of netrin receptor thrombospondin type 1 repeats by mammalian DPY19L1 and DPY19L3

2017 ◽  
Vol 114 (10) ◽  
pp. 2574-2579 ◽  
Author(s):  
Aleksandra Shcherbakova ◽  
Birgit Tiemann ◽  
Falk F. R. Buettner ◽  
Hans Bakker
Keyword(s):  
Endoplasmic Reticulum ◽  
Cell Surface ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Ovary Cells ◽  
The Third ◽  
The Stability ◽  
Thrombospondin Type 1 Repeats

Thrombospondin type 1 repeats (TSRs) occur in diverse proteins involved in adhesion and signaling. The two extracellular TSRs of the netrin receptor UNC5A contain WxxWxxWxxC motifs that can beC-mannosylated on all tryptophans. A singleC-mannosyltransferase (dumpy-19, DPY-19), modifying the first two tryptophans, occurs inCaenorhabditis elegans, but four putative enzymes (DPY-19–like 1–4, DPY19L1–4) exist in mammals. Single and triple CRISPR-Cas9 knockouts of the three homologs that are expressed in Chinese hamster ovary cells (DPY19L1, DPY19L3, and DPY19L4) and complementation experiments with mouse homologs showed that DPY19L1 preferentially mannosylates the first two tryptophans and DPY19L3 prefers the third, whereas DPY19L4 has no function in TSR glycosylation. Mannosylation by DPY19L1 but not DPY19L3 is required for transport of UNC5A from the endoplasmic reticulum to the cell surface. In vertebrates, a newC-mannosyltransferase has apparently evolved to increase glycosylation of TSRs, potentially to increase the stability of the structurally essential tryptophan ladder or to provide additional adhesion functions.

scholarly journals Reconstitution of Brefeldin A–induced Golgi Tubulation and Fusion with the Endoplasmic Reticulum in Semi-Intact Chinese Hamster Ovary Cells

2000 ◽  
Vol 11 (9) ◽  
pp. 3073-3087 ◽  
Author(s):  
Fumi Kano ◽  
Yasushi Sako ◽  
Mitsuo Tagaya ◽  
Toshio Yanagida ◽  
Masayuki Murata
Keyword(s):  
Endoplasmic Reticulum ◽  
Golgi Complex ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Brefeldin A ◽  
Tubule Formation ◽  
Intact Cells ◽  
Ovary Cells ◽  
Kinetics Of

The fungal metabolite brefeldin A (BFA) induces the disassembly of the Golgi complex in mammalian cells. The drug seems to accentuate tubule formation and causes the subsequent fusion with the endoplasmic reticulum (ER). To investigate the biochemical requirements and kinetics of BFA-induced Golgi disassembly, we have reconstituted the process of green fluorescent protein–tagged Golgi complex disassembly in streptolysin O–permeabilized semi-intact Chinese hamster ovary cells. For quantitative analysis of the morphological changes to the Golgi complex in semi-intact cells, we developed a novel morphometric analysis. Based on this analysis, we have dissected the BFA-induced Golgi disassembly process biochemically into two processes, Golgi tubule formation and fusion with the ER, and found that the formation is induced by only ATP and the residual factors in the cells and that the subsequent fusion is mediated in anN-ethylmaleimide–sensitive factor–dependent manner via Golgi tubules. Tubulation occurs by two pathways that depend on either microtubule integrity or exogenously added cytosol. In the presence of GTPγS, coat protein I inhibited the Golgi tubule fusion with the ER but showed no apparent effect on tubulation. Additionally, we analyzed the kinetics of tubulation and fusion independently in nocodazole-treated and -untreated semi-intact cells and found that tubulation is a rate-limiting step of the Golgi disassembly.

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