Proteomic Shifts Reflecting Oxidative Stress and Reduced Capacity for Protein Synthesis, and Alterations to Mitochondrial Membranes in <i>Neurospora crassa</i> Lacking VDAC
Voltage-dependent anion-selective channels (VDAC) maintain the bidirectional flow of small metabolites across the mitochondrial outer membrane and participate in the regulation of multiple cellular processes. To understand the roles of VDAC in cellular homeostasis, preliminary proteomic analyses of...
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MDPI AG
2022-01-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/10/2/198 |
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| author | Sabbir R. Shuvo Anna Motnenko Oleg V. Krokhin Victor Spicer Deborah A. Court |
| author_facet | Sabbir R. Shuvo Anna Motnenko Oleg V. Krokhin Victor Spicer Deborah A. Court |
| author_sort | Sabbir R. Shuvo |
| collection | DOAJ |
| description | Voltage-dependent anion-selective channels (VDAC) maintain the bidirectional flow of small metabolites across the mitochondrial outer membrane and participate in the regulation of multiple cellular processes. To understand the roles of VDAC in cellular homeostasis, preliminary proteomic analyses of S100 cytosolic and mitochondria-enriched fractions from a VDAC-less <i>Neurospora crassa</i> strain (ΔPor-1) were performed. In the variant cells, less abundant proteins include subunits of translation initiation factor eIF-2, enzymes in the shikimate pathway leading to precursors of aromatic amino acids, and enzymes involved in sulfate assimilation and in the synthesis of methionine, cysteine, alanine, serine, and threonine. In contrast, some of the more abundant proteins are involved in electron flow, such as the α subunit of the electron transfer flavoprotein and lactate dehydrogenase, which is involved in one pathway leading to pyruvate synthesis. Increased levels of catalase and catalase activity support predicted increased levels of oxidative stress in ΔPor-1 cells, and higher levels of protein disulfide isomerase suggest activation of the unfolded protein response in the endoplasmic reticulum. ΔPor-1 cells are cold-sensitive, which led us to investigate the impact of the absence of VDAC on several mitochondrial membrane characteristics. Mitochondrial membranes in ΔPor-1 are more fluid than those of wild-type cells, the ratio of C18:1 to C18:3n3 acyl chains is reduced, and ergosterol levels are lower. In summary, these initial results indicate that VDAC-less <i>N. crassa</i> cells are characterized by a lower abundance of proteins involved in amino acid and protein synthesis and by increases in some associated with pyruvate metabolism and stress responses. Membrane lipids and hyphal morphology are also impacted by the absence of VDAC. |
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| spelling | doaj.art-4e7543e918544387aff29726f24e3e0c2023-11-23T21:12:25ZengMDPI AGMicroorganisms2076-26072022-01-0110219810.3390/microorganisms10020198Proteomic Shifts Reflecting Oxidative Stress and Reduced Capacity for Protein Synthesis, and Alterations to Mitochondrial Membranes in <i>Neurospora crassa</i> Lacking VDACSabbir R. Shuvo0Anna Motnenko1Oleg V. Krokhin2Victor Spicer3Deborah A. Court4Department of Biochemistry and Microbiology, North South University, Dhaka 1229, BangladeshDepartment of Microbiology, University of Manitoba, Winnipeg, MB R3T 2N2, CanadaDepartment of Internal Medicine, University of Manitoba, Winnipeg, MB R3T 2N2, CanadaManitoba Centre for Proteomics and Systems Biology, Winnipeg, MB R3E 3P4, CanadaDepartment of Microbiology, University of Manitoba, Winnipeg, MB R3T 2N2, CanadaVoltage-dependent anion-selective channels (VDAC) maintain the bidirectional flow of small metabolites across the mitochondrial outer membrane and participate in the regulation of multiple cellular processes. To understand the roles of VDAC in cellular homeostasis, preliminary proteomic analyses of S100 cytosolic and mitochondria-enriched fractions from a VDAC-less <i>Neurospora crassa</i> strain (ΔPor-1) were performed. In the variant cells, less abundant proteins include subunits of translation initiation factor eIF-2, enzymes in the shikimate pathway leading to precursors of aromatic amino acids, and enzymes involved in sulfate assimilation and in the synthesis of methionine, cysteine, alanine, serine, and threonine. In contrast, some of the more abundant proteins are involved in electron flow, such as the α subunit of the electron transfer flavoprotein and lactate dehydrogenase, which is involved in one pathway leading to pyruvate synthesis. Increased levels of catalase and catalase activity support predicted increased levels of oxidative stress in ΔPor-1 cells, and higher levels of protein disulfide isomerase suggest activation of the unfolded protein response in the endoplasmic reticulum. ΔPor-1 cells are cold-sensitive, which led us to investigate the impact of the absence of VDAC on several mitochondrial membrane characteristics. Mitochondrial membranes in ΔPor-1 are more fluid than those of wild-type cells, the ratio of C18:1 to C18:3n3 acyl chains is reduced, and ergosterol levels are lower. In summary, these initial results indicate that VDAC-less <i>N. crassa</i> cells are characterized by a lower abundance of proteins involved in amino acid and protein synthesis and by increases in some associated with pyruvate metabolism and stress responses. Membrane lipids and hyphal morphology are also impacted by the absence of VDAC.https://www.mdpi.com/2076-2607/10/2/198VDAC<i>Neurospora crassa</i>mitochondriacytosolproteomicsunfolded protein response |
| spellingShingle | Sabbir R. Shuvo Anna Motnenko Oleg V. Krokhin Victor Spicer Deborah A. Court Proteomic Shifts Reflecting Oxidative Stress and Reduced Capacity for Protein Synthesis, and Alterations to Mitochondrial Membranes in <i>Neurospora crassa</i> Lacking VDAC Microorganisms VDAC <i>Neurospora crassa</i> mitochondria cytosol proteomics unfolded protein response |
| title | Proteomic Shifts Reflecting Oxidative Stress and Reduced Capacity for Protein Synthesis, and Alterations to Mitochondrial Membranes in <i>Neurospora crassa</i> Lacking VDAC |
| title_full | Proteomic Shifts Reflecting Oxidative Stress and Reduced Capacity for Protein Synthesis, and Alterations to Mitochondrial Membranes in <i>Neurospora crassa</i> Lacking VDAC |
| title_fullStr | Proteomic Shifts Reflecting Oxidative Stress and Reduced Capacity for Protein Synthesis, and Alterations to Mitochondrial Membranes in <i>Neurospora crassa</i> Lacking VDAC |
| title_full_unstemmed | Proteomic Shifts Reflecting Oxidative Stress and Reduced Capacity for Protein Synthesis, and Alterations to Mitochondrial Membranes in <i>Neurospora crassa</i> Lacking VDAC |
| title_short | Proteomic Shifts Reflecting Oxidative Stress and Reduced Capacity for Protein Synthesis, and Alterations to Mitochondrial Membranes in <i>Neurospora crassa</i> Lacking VDAC |
| title_sort | proteomic shifts reflecting oxidative stress and reduced capacity for protein synthesis and alterations to mitochondrial membranes in i neurospora crassa i lacking vdac |
| topic | VDAC <i>Neurospora crassa</i> mitochondria cytosol proteomics unfolded protein response |
| url | https://www.mdpi.com/2076-2607/10/2/198 |
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