title: Cloning and functional analysis of the dpm2 and dpm3 genes from Trichoderma reesei expressed in a Saccharomyces cerevisiae dpm1Δ mutant strain.
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.
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5a, 02-106 Warsaw, Poland.
authors.forename: Patrycja
authors.initials: P
authors.lastname: Zembek
authors.forename: Urszula
authors.initials: U
authors.lastname: Perlińska-Lenart
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Rawa
authors.forename: Wioletta
authors.initials: W
authors.lastname: Górka-Nieć
authors.forename: Grażyna
authors.initials: G
authors.lastname: Palamarczyk
authors.forename: Joanna S
authors.initials: JS
authors.lastname: Kruszewska
mesh_terms: D003001:Cloning, Molecular; D015870:Gene Expression; D005838:Genotype; D008364:Mannosyltransferases / Q000235:genetics* / Q000378:metabolism*; D009154:Mutation / Q000235:genetics*; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D014242:Trichoderma / Q000201:enzymology* / Q000235:genetics
chemical_list: D021122:Protein Subunits; D008364:Mannosyltransferases; C022755:dolichyl-phosphate beta-D-mannosyltransferase