Protein Glycosylation in<i>Aspergillus fumigatus</i>Is Essential for Cell Wall Synthesis and Serves as a Promising Model of Multicellular Eukaryotic Development

Jin, Cheng

doi:10.1155/2012/654251

Cited by 36 publications

(20 citation statements)

References 173 publications

(209 reference statements)

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“…Except N -linked glycans, we also observed some O -linked glycans from A. niger , which is consistent with the literature report that O - and N -linked glycans often coexist in complex biological system such as the fungus Aspergillus fumigatus (Jin, 2012). The direct proof of O -linked glycan can be provided by gHBMC (Fig.…”

Section: Resultssupporting

confidence: 92%

Structural analysis of N- and O-glycans using ZIC-HILIC/dialysis coupled to NMR detection

Feng

Deng

et al. 2014

Fungal Genetics and Biology

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Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor–ligand and cell–cell interaction, and plays a crucial role in many biological functions. However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. In this study, we developed a glycan enrichment strategy that couples Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) with dialysis to enrich the glycans from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate. Our results suggest that the ZIC-HILIC enrichment coupled with dialysis is a simple, fast, and efficient sample preparation approach. The approach was thus applied to analysis of a biological complex sample, the pronase E digest of the secreted proteins from the fungus Aspergillus niger. The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. In all, our study provides compelling evidence that ZIC-HILIC separation coupled with dialysis is very effective and accessible in preparing glycans for the downstream NMR analysis, which could greatly facilitate the future NMR-based glycoproteomics research.

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Section: Resultssupporting

confidence: 92%

Structural analysis of N- and O-glycans using ZIC-HILIC/dialysis coupled to NMR detection

Feng

Deng

et al. 2014

Fungal Genetics and Biology

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“…Many of these known secreted proteins are shown to first localize to the cell wall via the secretory pathway and eventually diffuse to the environment. In silico analysis of FmqD predicts that it bears attributes indicative of secretion through the classical ER‐Golgi‐mediated secretory pathway including an N‐terminal signal peptide and putative surface‐exposed N‐glycosylation sites that have been shown to be important in the proper folding and localization of other A. fumigatus cell wall proteins (Zhang et al ., 2008; 2009; Kotz et al ., ; Li et al ., ; Jin, ). Our study supports trafficking of FmqD through this pathway as deletion of the signal peptide abolished FmqD cell wall localization and greatly reduced FqC production [despite knowing that the signal peptide is not required for enzyme activity in vitro (Ames et al ., )].…”

Section: Discussionmentioning

confidence: 99%

Co-ordination between BrlA regulation and secretion of the oxidoreductase FmqD directs selective accumulation of fumiquinazoline C to conidial tissues inAspergillus fumigatus

et al. 2014

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SUMMARY Aerial spores, crucial for propagation and dispersal of the Kingdom Fungi, are commonly the initial inoculum of pathogenic fungi. Natural products (secondary metabolites) have been correlated with fungal spore development and enhanced virulence in the human pathogen Aspergillus fumigatus but mechanisms for metabolite deposition in the spore are unknown. Metabolomic profiling of A. fumigatus deletion mutants of fumiquinazoline (Fq) cluster genes reveal that the first two products of the Fq cluster, FqF and FqA, are produced to comparable levels in all fungal tissues but the final enzymatically-derived product, FqC, predominantly accumulates in the fungal spore. Loss of the sporulation-specific transcription factor, BrlA, yields a strain unable to produce FqA or FqC. Fluorescence microscopy showed FmqD, the oxidoreductase required to generate FqC, was secreted via the Golgi apparatus to the cell wall in an actin-dependent manner. In contrast, all other members of the Fq pathway including the putative transporter, FmqE – which had no effect on Fq biosynthesis – were internal to the hyphae. The coordination of BrlA-mediated tissue specificity with FmqD secretion to the cell wall presents a previously undescribed mechanism to direct localization of specific secondary metabolites to spores of the differentiating fungus.

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“…Finally, this structure is utilized as molecular scaffold by mannosyltransferases and galactosyltransferases to synthesize the N-linked glycan outer chain. Putative proteins participating in this step are TR_65646, TR_4561, TR_80340, TR_46443, TR_58609, TR_81211, TR_82551, TR_105557, TR_79832, TR_109361, TR_21576, TR_69868, TR_69211, TR_46421, TR_66687, and TR_48178 in T. reesei; TA_41037, TA_132346, TA_133591, TA_132601, TA_30858, TA_143140, TA_132930, TA_301583, TA_46185, TA_81367, TA_297159, TA_38402, TA_83451, TA_240794, TA_291006, and TA_301560 in T. atroviride; TV_66986, TV_32495, TV_89619, TV_231261, TV_87298, TV_82029, TV_83396, TV_184845, TV_77126, TV_31851, TV_76470, TV_33900, TV_73292, TV_83426, TV_183497, (549,560,561). These are enzymes classified within family 47 of the glycosylhydrolases (562), and along with the ER ␣1,2-mannosidase form the class I mannosidases (563).…”

Section: Glycosylationmentioning

confidence: 99%

“…11), where an oligosaccharide is covalently attached to Asn and Thr/ Ser residues, respectively, and the addition of GPI at the C terminus, which is known as a GPI anchor (548)(549)(550). These biosynthetic pathways are essential for cell viability, to maintain proper cell wall structure and organization, and for virulence (549)(550)(551).…”

Section: Glycosylationmentioning

confidence: 99%

“…Interestingly, T. reesei and T. atroviride, but not T. virens, contain a fourth gene that likely encodes a Golgi complex ␣1,2-mannosidase (TR_111953 and TA_155013), as they contain a Met and Leu at the equivalent position of Arg 273 . Other filamentous fungi analyzed so far do not have this extra mannosidase activity within the Golgi complex (549,563); thus, it is currently unknown how this enzyme could participate within the N-linked glycan elaboration.…”

Section: Glycosylationmentioning

confidence: 99%

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The Genomes of Three Uneven Siblings: Footprints of the Lifestyles of Three Trichoderma Species

Schmoll

Dattenböck

Carreras-Villaseñor

et al. 2016

Microbiol Mol Biol Rev

165

195

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SUMMARYThe genusTrichodermacontains fungi with high relevance for humans, with applications in enzyme production for plant cell wall degradation and use in biocontrol. Here, we provide a broad, comprehensive overview of the genomic content of these species for “hot topic” research aspects, including CAZymes, transport, transcription factors, and development, along with a detailed analysis and annotation of less-studied topics, such as signal transduction, genome integrity, chromatin, photobiology, or lipid, sulfur, and nitrogen metabolism inT. reesei,T. atroviride, andT. virens, and we open up new perspectives to those topics discussed previously. In total, we covered more than 2,000 of the predicted 9,000 to 11,000 genes of eachTrichodermaspecies discussed, which is >20% of the respective gene content. Additionally, we considered available transcriptome data for the annotated genes. Highlights of our analyses include overall carbohydrate cleavage preferences due to the different genomic contents and regulation of the respective genes. We found light regulation of many sulfur metabolic genes. Additionally, a new Golgi 1,2-mannosidase likely involved inN-linked glycosylation was detected, as were indications for the ability ofTrichodermaspp. to generate hybrid galactose-containingN-linked glycans. The genomic inventory of effector proteins revealed numerous compounds unique toTrichoderma, and these warrant further investigation. We found interesting expansions in theTrichodermagenus in several signaling pathways, such as G-protein-coupled receptors, RAS GTPases, and casein kinases. A particularly interesting feature absolutely unique toT. atrovirideis the duplication of the alternative sulfur amino acid synthesis pathway.

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Protein Glycosylation inAspergillus fumigatusIs Essential for Cell Wall Synthesis and Serves as a Promising Model of Multicellular Eukaryotic Development

Cited by 36 publications

References 173 publications

Structural analysis of N- and O-glycans using ZIC-HILIC/dialysis coupled to NMR detection

Structural analysis of N- and O-glycans using ZIC-HILIC/dialysis coupled to NMR detection

Co-ordination between BrlA regulation and secretion of the oxidoreductase FmqD directs selective accumulation of fumiquinazoline C to conidial tissues inAspergillus fumigatus

The Genomes of Three Uneven Siblings: Footprints of the Lifestyles of Three Trichoderma Species

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