Inositol Phosphoryl Transferase, Ipt1, Is a Critical Determinant of Azole Resistance and Virulence Phenotypes in <i>Candida glabrata</i>
In this study, we have specifically blocked a key step of sphingolipid (SL) biosynthesis in <i>Candida glabrata</i> by disruption of the orthologs of ScIpt1 and ScSkn1. Based on their close homology with <i>S. cerevisiae</i> counterparts, the proteins are predicted to catalyz...
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MDPI AG
2022-06-01
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| 丛编: | Journal of Fungi |
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| author | Garima Shahi Mohit Kumar Nitesh Kumar Khandelwal Atanu Banerjee Parijat Sarkar Sonam Kumari Brooke D. Esquivel Neeraj Chauhan Amitabha Chattopadhyay Theodore C. White Naseem A. Gaur Ashutosh Singh Rajendra Prasad |
| author_facet | Garima Shahi Mohit Kumar Nitesh Kumar Khandelwal Atanu Banerjee Parijat Sarkar Sonam Kumari Brooke D. Esquivel Neeraj Chauhan Amitabha Chattopadhyay Theodore C. White Naseem A. Gaur Ashutosh Singh Rajendra Prasad |
| author_sort | Garima Shahi |
| collection | DOAJ |
| description | In this study, we have specifically blocked a key step of sphingolipid (SL) biosynthesis in <i>Candida glabrata</i> by disruption of the orthologs of ScIpt1 and ScSkn1. Based on their close homology with <i>S. cerevisiae</i> counterparts, the proteins are predicted to catalyze the addition of a phosphorylinositol group onto mannosyl inositolphosphoryl ceramide (MIPC) to form mannosyl diinositolphosphoryl ceramide (M(IP)<sub>2</sub>C), which accounts for the majority of complex SL structures in <i>S. cerevisiae</i> membranes. High throughput lipidome analysis confirmed the accumulation of MIPC structures in <i>ΔCgipt1</i> and <i>ΔCgskn1</i> cells, albeit to lesser extent in the latter. Noticeably, <i>ΔCgipt1</i> cells showed an increased susceptibility to azoles; however, <i>ΔCgskn1</i> cells showed no significant changes in the drug susceptibility profiles. Interestingly, the azole susceptible phenotype of <i>ΔCgipt1</i> cells seems to be independent of the ergosterol content. <i>ΔCgipt1</i> cells displayed altered lipid homeostasis, increased membrane fluidity as well as high diffusion of radiolabeled fluconazole (<sup>3</sup>H-FLC), which could together influence the azole susceptibility of <i>C. glabrata</i>. Furthermore, in vivo experiments also confirmed compromised virulence of the <i>ΔCgipt1</i> strain. Contrarily, specific functions of CgSkn1 remain unclear. |
| first_indexed | 2024-03-09T10:15:59Z |
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| id | doaj.art-042d099b74d3401890bb9f48bc56b6e7 |
| institution | Directory Open Access Journal |
| issn | 2309-608X |
| language | English |
| last_indexed | 2024-03-09T10:15:59Z |
| publishDate | 2022-06-01 |
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| series | Journal of Fungi |
| spelling | doaj.art-042d099b74d3401890bb9f48bc56b6e72023-12-01T22:19:46ZengMDPI AGJournal of Fungi2309-608X2022-06-018765110.3390/jof8070651Inositol Phosphoryl Transferase, Ipt1, Is a Critical Determinant of Azole Resistance and Virulence Phenotypes in <i>Candida glabrata</i>Garima Shahi0Mohit Kumar1Nitesh Kumar Khandelwal2Atanu Banerjee3Parijat Sarkar4Sonam Kumari5Brooke D. Esquivel6Neeraj Chauhan7Amitabha Chattopadhyay8Theodore C. White9Naseem A. Gaur10Ashutosh Singh11Rajendra Prasad12Amity Institute of Biotechnology and Integrative Science and Health, Amity University Gurgaon, Gurgaon 122412, IndiaAmity Institute of Biotechnology and Integrative Science and Health, Amity University Gurgaon, Gurgaon 122412, IndiaDepartment of Chemistry and Biochemistry, University of Arizona, Tucson, AZ 85721, USAAmity Institute of Biotechnology and Integrative Science and Health, Amity University Gurgaon, Gurgaon 122412, IndiaCSIR-Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad 500007, IndiaYeast Biofuel Group, International Centre for Genetic Engineering and Biotechnology, New Delhi 110067, IndiaSchool of Biological and Chemical Sciences, University of Missouri at Kansas City, Kansas City, MO 64110, USADepartment of Microbiology, Biochemistry and Molecular Genetics, New Jersey Medical School, Rutgers, The State University of New Jersey, Newark, NJ 07103, USACSIR-Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad 500007, IndiaSchool of Biological and Chemical Sciences, University of Missouri at Kansas City, Kansas City, MO 64110, USAYeast Biofuel Group, International Centre for Genetic Engineering and Biotechnology, New Delhi 110067, IndiaDepartment of Biochemistry, University of Lucknow, Lucknow 226007, IndiaAmity Institute of Biotechnology and Integrative Science and Health, Amity University Gurgaon, Gurgaon 122412, IndiaIn this study, we have specifically blocked a key step of sphingolipid (SL) biosynthesis in <i>Candida glabrata</i> by disruption of the orthologs of ScIpt1 and ScSkn1. Based on their close homology with <i>S. cerevisiae</i> counterparts, the proteins are predicted to catalyze the addition of a phosphorylinositol group onto mannosyl inositolphosphoryl ceramide (MIPC) to form mannosyl diinositolphosphoryl ceramide (M(IP)<sub>2</sub>C), which accounts for the majority of complex SL structures in <i>S. cerevisiae</i> membranes. High throughput lipidome analysis confirmed the accumulation of MIPC structures in <i>ΔCgipt1</i> and <i>ΔCgskn1</i> cells, albeit to lesser extent in the latter. Noticeably, <i>ΔCgipt1</i> cells showed an increased susceptibility to azoles; however, <i>ΔCgskn1</i> cells showed no significant changes in the drug susceptibility profiles. Interestingly, the azole susceptible phenotype of <i>ΔCgipt1</i> cells seems to be independent of the ergosterol content. <i>ΔCgipt1</i> cells displayed altered lipid homeostasis, increased membrane fluidity as well as high diffusion of radiolabeled fluconazole (<sup>3</sup>H-FLC), which could together influence the azole susceptibility of <i>C. glabrata</i>. Furthermore, in vivo experiments also confirmed compromised virulence of the <i>ΔCgipt1</i> strain. Contrarily, specific functions of CgSkn1 remain unclear.https://www.mdpi.com/2309-608X/8/7/651<i>Candida glabrata</i>sphingolipidsinositolphosphorylceramidelipidomicsdrug resistancevirulence |
| spellingShingle | Garima Shahi Mohit Kumar Nitesh Kumar Khandelwal Atanu Banerjee Parijat Sarkar Sonam Kumari Brooke D. Esquivel Neeraj Chauhan Amitabha Chattopadhyay Theodore C. White Naseem A. Gaur Ashutosh Singh Rajendra Prasad Inositol Phosphoryl Transferase, Ipt1, Is a Critical Determinant of Azole Resistance and Virulence Phenotypes in <i>Candida glabrata</i> Journal of Fungi <i>Candida glabrata</i> sphingolipids inositolphosphorylceramide lipidomics drug resistance virulence |
| title | Inositol Phosphoryl Transferase, Ipt1, Is a Critical Determinant of Azole Resistance and Virulence Phenotypes in <i>Candida glabrata</i> |
| title_full | Inositol Phosphoryl Transferase, Ipt1, Is a Critical Determinant of Azole Resistance and Virulence Phenotypes in <i>Candida glabrata</i> |
| title_fullStr | Inositol Phosphoryl Transferase, Ipt1, Is a Critical Determinant of Azole Resistance and Virulence Phenotypes in <i>Candida glabrata</i> |
| title_full_unstemmed | Inositol Phosphoryl Transferase, Ipt1, Is a Critical Determinant of Azole Resistance and Virulence Phenotypes in <i>Candida glabrata</i> |
| title_short | Inositol Phosphoryl Transferase, Ipt1, Is a Critical Determinant of Azole Resistance and Virulence Phenotypes in <i>Candida glabrata</i> |
| title_sort | inositol phosphoryl transferase ipt1 is a critical determinant of azole resistance and virulence phenotypes in i candida glabrata i |
| topic | <i>Candida glabrata</i> sphingolipids inositolphosphorylceramide lipidomics drug resistance virulence |
| url | https://www.mdpi.com/2309-608X/8/7/651 |
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