Characterization of PcSTT3B as a Key Oligosaccharyltransferase Subunit Involved in N-glycosylation and Its Role in Development and Pathogenicity of <i>Phytophthora capsici</i>
Asparagine (Asn, N)-linked glycosylation is a conserved process and an essential post-translational modification that occurs on the NXT/S motif of the nascent polypeptides in endoplasmic reticulum (ER). The mechanism of N-glycosylation and biological functions of key catalytic enzymes involved in th...
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MDPI AG
2023-04-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/8/7500 |
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| author | Tongshan Cui Quanhe Ma Fan Zhang Shanshan Chen Can Zhang Jianjun Hao Xili Liu |
| author_facet | Tongshan Cui Quanhe Ma Fan Zhang Shanshan Chen Can Zhang Jianjun Hao Xili Liu |
| author_sort | Tongshan Cui |
| collection | DOAJ |
| description | Asparagine (Asn, N)-linked glycosylation is a conserved process and an essential post-translational modification that occurs on the NXT/S motif of the nascent polypeptides in endoplasmic reticulum (ER). The mechanism of N-glycosylation and biological functions of key catalytic enzymes involved in this process are rarely documented for oomycetes. In this study, an N-glycosylation inhibitor tunicamycin (TM) hampered the mycelial growth, sporangial release, and zoospore production of <i>Phytophthora capsici</i>, indicating that N-glycosylation was crucial for oomycete growth development. Among the key catalytic enzymes involved in N-glycosylation, the <i>PcSTT3B</i> gene was characterized by its functions in <i>P. capsici</i>. As a core subunit of the oligosaccharyltransferase (OST) complex, the staurosporine and temperature sensive 3B (STT3B) subunit were critical for the catalytic activity of OST. The <i>PcSTT3B</i> gene has catalytic activity and is highly conservative in <i>P. capsici</i>. By using a CRISPR/Cas9-mediated gene replacement system to delete the <i>PcSTT3B</i> gene, the transformants impaired mycelial growth, sporangial release, zoospore production, and virulence. The <i>PcSTT3B</i>-deleted transformants were more sensitive to an ER stress inducer TM and display low glycoprotein content in the mycelia, suggesting that PcSTT3B was associated with ER stress responses and N-glycosylation. Therefore, PcSTT3B was involved in the development, pathogenicity, and N-glycosylation of <i>P. capsici</i>. |
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| publishDate | 2023-04-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-dda4581209f54561a5b0a4486ee384192023-11-17T19:41:23ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-04-01248750010.3390/ijms24087500Characterization of PcSTT3B as a Key Oligosaccharyltransferase Subunit Involved in N-glycosylation and Its Role in Development and Pathogenicity of <i>Phytophthora capsici</i>Tongshan Cui0Quanhe Ma1Fan Zhang2Shanshan Chen3Can Zhang4Jianjun Hao5Xili Liu6Department of Plant Pathology, College of Plant Protection, China Agricultural University, Beijing 100193, ChinaDepartment of Plant Pathology, College of Plant Protection, China Agricultural University, Beijing 100193, ChinaDepartment of Plant Pathology, College of Plant Protection, China Agricultural University, Beijing 100193, ChinaDepartment of Plant Pathology, College of Plant Protection, China Agricultural University, Beijing 100193, ChinaDepartment of Plant Pathology, College of Plant Protection, China Agricultural University, Beijing 100193, ChinaSchool of Food and Agriculture, University of Maine, Orono, ME 04469, USADepartment of Plant Pathology, College of Plant Protection, China Agricultural University, Beijing 100193, ChinaAsparagine (Asn, N)-linked glycosylation is a conserved process and an essential post-translational modification that occurs on the NXT/S motif of the nascent polypeptides in endoplasmic reticulum (ER). The mechanism of N-glycosylation and biological functions of key catalytic enzymes involved in this process are rarely documented for oomycetes. In this study, an N-glycosylation inhibitor tunicamycin (TM) hampered the mycelial growth, sporangial release, and zoospore production of <i>Phytophthora capsici</i>, indicating that N-glycosylation was crucial for oomycete growth development. Among the key catalytic enzymes involved in N-glycosylation, the <i>PcSTT3B</i> gene was characterized by its functions in <i>P. capsici</i>. As a core subunit of the oligosaccharyltransferase (OST) complex, the staurosporine and temperature sensive 3B (STT3B) subunit were critical for the catalytic activity of OST. The <i>PcSTT3B</i> gene has catalytic activity and is highly conservative in <i>P. capsici</i>. By using a CRISPR/Cas9-mediated gene replacement system to delete the <i>PcSTT3B</i> gene, the transformants impaired mycelial growth, sporangial release, zoospore production, and virulence. The <i>PcSTT3B</i>-deleted transformants were more sensitive to an ER stress inducer TM and display low glycoprotein content in the mycelia, suggesting that PcSTT3B was associated with ER stress responses and N-glycosylation. Therefore, PcSTT3B was involved in the development, pathogenicity, and N-glycosylation of <i>P. capsici</i>.https://www.mdpi.com/1422-0067/24/8/7500N-glycosylationoligosaccharyltransferasePcSTT3Bvirulence<i>Phytophthora capsici</i> |
| spellingShingle | Tongshan Cui Quanhe Ma Fan Zhang Shanshan Chen Can Zhang Jianjun Hao Xili Liu Characterization of PcSTT3B as a Key Oligosaccharyltransferase Subunit Involved in N-glycosylation and Its Role in Development and Pathogenicity of <i>Phytophthora capsici</i> International Journal of Molecular Sciences N-glycosylation oligosaccharyltransferase PcSTT3B virulence <i>Phytophthora capsici</i> |
| title | Characterization of PcSTT3B as a Key Oligosaccharyltransferase Subunit Involved in N-glycosylation and Its Role in Development and Pathogenicity of <i>Phytophthora capsici</i> |
| title_full | Characterization of PcSTT3B as a Key Oligosaccharyltransferase Subunit Involved in N-glycosylation and Its Role in Development and Pathogenicity of <i>Phytophthora capsici</i> |
| title_fullStr | Characterization of PcSTT3B as a Key Oligosaccharyltransferase Subunit Involved in N-glycosylation and Its Role in Development and Pathogenicity of <i>Phytophthora capsici</i> |
| title_full_unstemmed | Characterization of PcSTT3B as a Key Oligosaccharyltransferase Subunit Involved in N-glycosylation and Its Role in Development and Pathogenicity of <i>Phytophthora capsici</i> |
| title_short | Characterization of PcSTT3B as a Key Oligosaccharyltransferase Subunit Involved in N-glycosylation and Its Role in Development and Pathogenicity of <i>Phytophthora capsici</i> |
| title_sort | characterization of pcstt3b as a key oligosaccharyltransferase subunit involved in n glycosylation and its role in development and pathogenicity of i phytophthora capsici i |
| topic | N-glycosylation oligosaccharyltransferase PcSTT3B virulence <i>Phytophthora capsici</i> |
| url | https://www.mdpi.com/1422-0067/24/8/7500 |
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