Abstract
The N-linked glycosylation is a ubiquitous protein modification in eukaryotic cells. During the N-linked glycan synthesis, the precursor Glc(3)Man(9)GlcNAc(2) is processed by endoplasmic reticulum (ER) glucosidases I, II and α1,2-mannosidase, before transporting to the Golgi complex for further structure modifications. In fungi of medical relevance, as Candida albicans and Aspergillus, it is well known that ER glycosidases are important for cell fitness, cell wall organization, virulence, and interaction with the immune system. Despite this, little is known about these enzymes in Sporothrix schenckii, the causative agent of human sporotrichosis. This limited knowledge is due in part to the lack of a genome sequence of this organism. In this work we used degenerate primers and inverse PCR approaches to isolate the open reading frame of S. schenckii ROT2, the encoding gene for α subunit of ER glucosidase II. This S. schenckii gene complemented a Saccharomyces cerevisiae rot2Δ mutant; however, when expressed in a C. albicans rot2Δ mutant, S. schenckii Rot2 partially increased the levels of α-glucosidase activity, but failed to restore the N-linked glycosylation defect associated to the mutation. To our knowledge, this is the first report where a gene involved in protein N-linked glycosylation is isolated from S. schenckii.
| Original language | English |
|---|---|
| Pages (from-to) | 910-918 |
| Number of pages | 9 |
| Journal | Fungal Biology |
| Volume | 116 |
| Issue number | 8 |
| DOIs | |
| Publication status | Published - Aug 2012 |
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Keywords
- amino acid sequence
- endoplasmic reticulum
- fungal proteins
- glycosylation
- humans
- molecular sequence data
- sequence alignment
- sporothrix
- alpha-glucosidases
Cite this
Isolation and functional characterization of Sporothrix schenckii ROT2, the encoding gene for the endoplasmic reticulum glucosidase II. / Robledo-Ortiz, Claudia I; Flores-Carreón, Arturo; Hernández-Cervantes, Arturo; Álvarez-Vargas, Aurelio; Lee, Keunsook K; Díaz-Jiménez, Diana F; Munro, Carol A; Cano-Canchola, Carmen; Mora-Montes, Héctor M.
In: Fungal Biology, Vol. 116, No. 8, 08.2012, p. 910-918.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Isolation and functional characterization of Sporothrix schenckii ROT2, the encoding gene for the endoplasmic reticulum glucosidase II
AU - Robledo-Ortiz, Claudia I
AU - Flores-Carreón, Arturo
AU - Hernández-Cervantes, Arturo
AU - Álvarez-Vargas, Aurelio
AU - Lee, Keunsook K
AU - Díaz-Jiménez, Diana F
AU - Munro, Carol A
AU - Cano-Canchola, Carmen
AU - Mora-Montes, Héctor M
N1 - Copyright © 2012 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.
PY - 2012/8
Y1 - 2012/8
N2 - The N-linked glycosylation is a ubiquitous protein modification in eukaryotic cells. During the N-linked glycan synthesis, the precursor Glc(3)Man(9)GlcNAc(2) is processed by endoplasmic reticulum (ER) glucosidases I, II and α1,2-mannosidase, before transporting to the Golgi complex for further structure modifications. In fungi of medical relevance, as Candida albicans and Aspergillus, it is well known that ER glycosidases are important for cell fitness, cell wall organization, virulence, and interaction with the immune system. Despite this, little is known about these enzymes in Sporothrix schenckii, the causative agent of human sporotrichosis. This limited knowledge is due in part to the lack of a genome sequence of this organism. In this work we used degenerate primers and inverse PCR approaches to isolate the open reading frame of S. schenckii ROT2, the encoding gene for α subunit of ER glucosidase II. This S. schenckii gene complemented a Saccharomyces cerevisiae rot2Δ mutant; however, when expressed in a C. albicans rot2Δ mutant, S. schenckii Rot2 partially increased the levels of α-glucosidase activity, but failed to restore the N-linked glycosylation defect associated to the mutation. To our knowledge, this is the first report where a gene involved in protein N-linked glycosylation is isolated from S. schenckii.
AB - The N-linked glycosylation is a ubiquitous protein modification in eukaryotic cells. During the N-linked glycan synthesis, the precursor Glc(3)Man(9)GlcNAc(2) is processed by endoplasmic reticulum (ER) glucosidases I, II and α1,2-mannosidase, before transporting to the Golgi complex for further structure modifications. In fungi of medical relevance, as Candida albicans and Aspergillus, it is well known that ER glycosidases are important for cell fitness, cell wall organization, virulence, and interaction with the immune system. Despite this, little is known about these enzymes in Sporothrix schenckii, the causative agent of human sporotrichosis. This limited knowledge is due in part to the lack of a genome sequence of this organism. In this work we used degenerate primers and inverse PCR approaches to isolate the open reading frame of S. schenckii ROT2, the encoding gene for α subunit of ER glucosidase II. This S. schenckii gene complemented a Saccharomyces cerevisiae rot2Δ mutant; however, when expressed in a C. albicans rot2Δ mutant, S. schenckii Rot2 partially increased the levels of α-glucosidase activity, but failed to restore the N-linked glycosylation defect associated to the mutation. To our knowledge, this is the first report where a gene involved in protein N-linked glycosylation is isolated from S. schenckii.
KW - amino acid sequence
KW - endoplasmic reticulum
KW - fungal proteins
KW - glycosylation
KW - humans
KW - molecular sequence data
KW - sequence alignment
KW - sporothrix
KW - alpha-glucosidases
U2 - 10.1016/j.funbio.2012.06.002
DO - 10.1016/j.funbio.2012.06.002
M3 - Article
VL - 116
SP - 910
EP - 918
JO - Fungal Biology
JF - Fungal Biology
SN - 1878-6146
IS - 8
ER -