Protein O-mannosylation affects protein secretion, cell wall integrity and morphogenesis in Trichoderma reesei

Fungal Genet Biol. 2020 Nov;144:103440. doi: 10.1016/j.fgb.2020.103440. Epub 2020 Aug 3.


Protein O-mannosyltransferases (PMTs) initiate O-mannosylation of proteins in the ER. Trichoderma reesei strains displayed a single representative of each PMT subfamily, Trpmt1, Trpmt2 and Trpmt4. In this work, two knockout strains ΔTrpmt1and ΔTrpmt4were obtained. Both mutants showed retarded growth, defective cell walls, reduced conidiation and decreased protein secretion. Additionally, the ΔTrpmt1strain displayed a thermosensitive growth phenotype, while the ΔTrpmt4 strain showed abnormal polarity. Meanwhile, OETrpmt2 strain, in which the Trpmt2 was over-expressed, exhibited increased conidiation, enhanced protein secretion and abnormal polarity. Using a lectin enrichment method and MS/MS analysis, 173 O-glycoproteins, 295 O-glycopeptides and 649 O-mannosylation sites were identified as the targets of PMTs in T. reesei. These identified O-mannoproteins are involved in various physiological processes such as protein folding, sorting, transport, quality control and secretion, as well as cell wall integrity and polarity. By comparing proteins identified in the mutants and its parent strain, the potential specific protein substrates of PMTs were identified. Based on our results, TrPMT1 is specifically involved inO-mannosylation of intracellular soluble proteins and secreted proteins, specially glycosidases. TrPMT2 is involved inO-mannosylation of secreted proteins and GPI-anchor proteins, and TrPMT4 mainly modifies multiple transmembrane proteins. The TrPMT1-TrPMT4 complex is responsible for O-mannosylation of proteins involved in cell wall integrity. Overexpression of TrPMT2 enhances protein secretion, which might be a new strategy to improve expression efficiency in T. reesei.

Keywords: Glycopeptide; O-mannosylation; Protein O-mannosyltransferase; Protein secretion; Trichoderma reesei.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Wall / genetics
  • Fungal Proteins / biosynthesis*
  • Fungal Proteins / genetics
  • Glycosylation
  • Hypocreales / enzymology
  • Hypocreales / genetics*
  • Mannosyltransferases / genetics*
  • Morphogenesis / genetics*
  • Phenotype
  • Protein Transport / genetics
  • Tandem Mass Spectrometry


  • Fungal Proteins
  • Mannosyltransferases
  • protein O-mannosyltransferase

Supplementary concepts

  • Trichoderma reesei