Functional Analysis of an Essential GSP1/Ran Ortholog Gene, <i>CpRan1</i>, from the Chestnut Blight Fungus <i>Cryphonectria parasitica</i> Using a Heterokaryon
Functional analysis of a GSP1/Ran ortholog, <i>CpRan1</i>, from <i>Cryphonectria parasitica</i> was conducted. Genotype analysis revealed that the putative <i>CpRan1</i>-null mutant was a heterokaryotic transformant harboring two different types of nuclei, one wit...
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MDPI AG
2021-04-01
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| author | Yo-Han Ko Jeesun Chun Dae-Hyuk Kim |
| author_facet | Yo-Han Ko Jeesun Chun Dae-Hyuk Kim |
| author_sort | Yo-Han Ko |
| collection | DOAJ |
| description | Functional analysis of a GSP1/Ran ortholog, <i>CpRan1</i>, from <i>Cryphonectria parasitica</i> was conducted. Genotype analysis revealed that the putative <i>CpRan1</i>-null mutant was a heterokaryotic transformant harboring two different types of nuclei, one with the wild-type <i>CpRan1</i> allele and the other with the <i>CpRan1</i>-null mutant allele. The mycelial growth and colony morphology of the heterokaryotic transformant was normal. Microscopic analysis of the resulting conidia (aseptate and monokaryotic asexual spores) demonstrated that although normal germinating spores were observed from conidia harboring a nucleus with the wild-type <i>CpRan1</i> allele, a number of residual conidia that did not germinate existed. Complementation analysis using protoplasts from the heterokaryon with the wild-type <i>CpRan1</i> allele confirmed that the <i>CpRan1</i> gene is essential to <i>C. parasitica</i>. Complementation analysis using the various <i>CpRan1</i> chimera constructs allowed us to perform a functional analysis of essential amino acids of the <i>CpRan1</i>. Among the four suggested essential amino acids, Lys-97 for ubiquitination was determined to not be an essential residue. Moreover, the <i>CpRan1</i>-null mutant allele was successfully complemented with mouse Ran gene, which suggested that the biological function of Ran gene is evolutionary conserved and that our heterokaryon rescue can be applied for the functional analysis of heterologous genes. |
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| language | English |
| last_indexed | 2024-03-10T11:58:45Z |
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| spelling | doaj.art-394f1f5493ed4f548b1b9f98a434277a2023-11-21T17:06:23ZengMDPI AGJournal of Fungi2309-608X2021-04-017533210.3390/jof7050332Functional Analysis of an Essential GSP1/Ran Ortholog Gene, <i>CpRan1</i>, from the Chestnut Blight Fungus <i>Cryphonectria parasitica</i> Using a HeterokaryonYo-Han Ko0Jeesun Chun1Dae-Hyuk Kim2Department of Molecular Biology, Institute for Molecular Biology and Genetics, Jeonbuk National University, 567 Baekje-daero, Jeonju, Chonbuk 54896, KoreaDepartment of Molecular Biology, Institute for Molecular Biology and Genetics, Jeonbuk National University, 567 Baekje-daero, Jeonju, Chonbuk 54896, KoreaDepartment of Molecular Biology, Institute for Molecular Biology and Genetics, Jeonbuk National University, 567 Baekje-daero, Jeonju, Chonbuk 54896, KoreaFunctional analysis of a GSP1/Ran ortholog, <i>CpRan1</i>, from <i>Cryphonectria parasitica</i> was conducted. Genotype analysis revealed that the putative <i>CpRan1</i>-null mutant was a heterokaryotic transformant harboring two different types of nuclei, one with the wild-type <i>CpRan1</i> allele and the other with the <i>CpRan1</i>-null mutant allele. The mycelial growth and colony morphology of the heterokaryotic transformant was normal. Microscopic analysis of the resulting conidia (aseptate and monokaryotic asexual spores) demonstrated that although normal germinating spores were observed from conidia harboring a nucleus with the wild-type <i>CpRan1</i> allele, a number of residual conidia that did not germinate existed. Complementation analysis using protoplasts from the heterokaryon with the wild-type <i>CpRan1</i> allele confirmed that the <i>CpRan1</i> gene is essential to <i>C. parasitica</i>. Complementation analysis using the various <i>CpRan1</i> chimera constructs allowed us to perform a functional analysis of essential amino acids of the <i>CpRan1</i>. Among the four suggested essential amino acids, Lys-97 for ubiquitination was determined to not be an essential residue. Moreover, the <i>CpRan1</i>-null mutant allele was successfully complemented with mouse Ran gene, which suggested that the biological function of Ran gene is evolutionary conserved and that our heterokaryon rescue can be applied for the functional analysis of heterologous genes.https://www.mdpi.com/2309-608X/7/5/332<i>Cryphonectria parasitica</i>Ranheterokaryonhypovirulence |
| spellingShingle | Yo-Han Ko Jeesun Chun Dae-Hyuk Kim Functional Analysis of an Essential GSP1/Ran Ortholog Gene, <i>CpRan1</i>, from the Chestnut Blight Fungus <i>Cryphonectria parasitica</i> Using a Heterokaryon Journal of Fungi <i>Cryphonectria parasitica</i> Ran heterokaryon hypovirulence |
| title | Functional Analysis of an Essential GSP1/Ran Ortholog Gene, <i>CpRan1</i>, from the Chestnut Blight Fungus <i>Cryphonectria parasitica</i> Using a Heterokaryon |
| title_full | Functional Analysis of an Essential GSP1/Ran Ortholog Gene, <i>CpRan1</i>, from the Chestnut Blight Fungus <i>Cryphonectria parasitica</i> Using a Heterokaryon |
| title_fullStr | Functional Analysis of an Essential GSP1/Ran Ortholog Gene, <i>CpRan1</i>, from the Chestnut Blight Fungus <i>Cryphonectria parasitica</i> Using a Heterokaryon |
| title_full_unstemmed | Functional Analysis of an Essential GSP1/Ran Ortholog Gene, <i>CpRan1</i>, from the Chestnut Blight Fungus <i>Cryphonectria parasitica</i> Using a Heterokaryon |
| title_short | Functional Analysis of an Essential GSP1/Ran Ortholog Gene, <i>CpRan1</i>, from the Chestnut Blight Fungus <i>Cryphonectria parasitica</i> Using a Heterokaryon |
| title_sort | functional analysis of an essential gsp1 ran ortholog gene i cpran1 i from the chestnut blight fungus i cryphonectria parasitica i using a heterokaryon |
| topic | <i>Cryphonectria parasitica</i> Ran heterokaryon hypovirulence |
| url | https://www.mdpi.com/2309-608X/7/5/332 |
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