Cloning and functional analysis of the dpm2 and dpm3 genes from Trichoderma reesei expressed in a Saccharomyces cerevisiae dpm1Δ mutant strain
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Patrycja Zembek
Abstract
In Trichoderma reesei, dolichyl phosphate mannose (dpm) synthase, a key enzyme in the O-glycosylation process, requires three proteins for full activity. In this study, the dpm2 and dpm3 genes coding for the DPMII and DPMIII subunits of T. reesei DPM synthase were cloned and functionally analyzed after expression in the Saccharomyces cerevisiae dpm1Δ [genotype (BY4743; his3Δ1; /leu2Δ0; lys2Δ0; /ura3Δ0; YPR183w::kanMX4] mutant. It was found that apart from the catalytic subunit DPMI, the DPMIII subunit is also essential to form an active DPM synthase in yeast. Additional expression of the DPMII protein, considered to be a regulatory subunit of DPM synthase, decreased the enzymatic activity. We also characterized S. cerevisiae strains expressing the dpm1, 2, 3 or dpm1, 3 genes and analyzed the consequences of dpm expression on protein O-glycosylation in vivo and on the cell wall composition.
©2011 by Walter de Gruyter Berlin New York
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Articles in the same Issue
- Publisher’s Note
- Publisher’s Note
- GENES AND NUCLEIC ACIDS
- Intermolecular interaction between a branching ribozyme and associated homing endonuclease mRNA
- Type V collagen-induced upregulation of capn2 (large subunit of m-calpain) gene expression and DNA fragmentation in 8701-BC breast cancer cells
- PROTEIN STRUCTURE AND FUNCTION
- High catalytic efficiency and resistance to denaturing in bacterial Rho GTPase-activating proteins
- MEMBRANES, LIPIDS, GLYCOBIOLOGY
- Cloning and functional analysis of the dpm2 and dpm3 genes from Trichoderma reesei expressed in a Saccharomyces cerevisiae dpm1Δ mutant strain
- MOLECULAR MEDICINE
- Increased expression of the multidrug resistance-associated protein 1 (MRP1) in kidney glomeruli of streptozotocin-induced diabetic rats
- CELL BIOLOGY AND SIGNALING
- Investigating the effects of physiological bile acids on GLP-1 secretion and glucose tolerance in normal and GLP-1R-/- mice
- PROTEOLYSIS
- Angiotensin I-converting enzyme (ACE) activity and expression in rat central nervous system after sleep deprivation
- Cathepsins S, B and L with aminopeptidases display β-secretase activity associated with the pathogenesis of Alzheimer’s disease
- The role of cathepsin E in terminal differentiation of keratinocytes
- Poliovirus 3C proteinase inhibition by organotelluranes
