Identification of HUGT1 as a potential BiP activator and a cellular target for improvement of recombinant protein production using a cDNA screening system.
The development of a high-throughput functional genomic screening provides a novel and expeditious approach in identifying critical genes involved in specific biological processes. Here we describe a cell-based cDNA screening system to identify the transcription activators of BiP, an endoplasmic reticulum (ER) chaperone protein. BiP promoter contains the ER stress element which is commonly present in the genes involved in unfolded protein response (UPR) that regulates protein secretion in cells. Therefore, the positive regulators of BiP may also be utilized to improve the recombinant protein production through modulation of UPR. Four BiP activators, including human UDP-glucose:glycoprotein glucosyltransferase 1 (HUGT1), are identified by the cDNA screening. Overexpression of HUGT1 leads to a significant increase in the production of recombinant erythropoietin, interferon gamma, and monoclonal antibody in HEK293 cells. Our results demonstrate that the cDNA screening for BiP activators may be effective to identify the novel BiP regulators and HUGT1 may serve as an ideal target gene for improving the recombinant protein production in mammalian cells.
Ku
Sebastian Chih Yuan
SC
Expression Engineering Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), 20 Biopolis Way, #06-01, 138668 Singapore, Singapore.
Lwa
Teng Rhui
TR
Giam
Maybelline
M
Yap
Miranda Gek Sim
MG
Chao
Sheng-Hao
SH
D000911:Antibodies, Monoclonal / Q000235:genetics / Q000378:metabolism*; D001709:Biotechnology; D002460:Cell Line; D004721:Endoplasmic Reticulum; D000091342:Endoplasmic Reticulum Chaperone BiP; D004921:Erythropoietin / Q000235:genetics / Q000378:metabolism*; D015723:Gene Library; D018390:Gene Targeting / Q000379:methods / Q000639:trends; D005818:Genetic Engineering / Q000639:trends; D006360:Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007371:Interferon-gamma / Q000235:genetics / Q000378:metabolism*; D009004:Monosaccharide Transport Proteins / Q000235:genetics* / Q000378:metabolism; D017510:Protein Folding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D015533:Transcriptional Activation; D000099237:UDP-Galactose Translocators
D000911:Antibodies, Monoclonal; D000091342:Endoplasmic Reticulum Chaperone BiP; D004921:Erythropoietin; D006360:Heat-Shock Proteins; D007371:Interferon-gamma; D009004:Monosaccharide Transport Proteins; D011994:Recombinant Proteins; D000099237:UDP-Galactose Translocators