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Wsv156 targets mitochondria via Cq TOMM70 and triggers mito-aggresome formation. ( A ) Wsv156 expression increased following WSSV infection. Temporal expression of wsv156 at the mRNA (left panel) and protein level (right panel) was assessed by RT-qPCR and western blot, respectively, in Hpt cells after WSSV infection. ( B ) Wsv156 localized to mitochondria. Endogenous wsv156 protein in Hpt cells (left panel) and overexpressed wsv156 in HEK 293T cells (right panel) both predominantly co-fractionated with mitochondria. ( C ) Identification of host mitochondrial proteins interacting with wsv156 by affinity purification-mass spectrometry (AP-MS). ( D ) Wsv156 interacted with Cq TOMM70. Co-IP <t>with</t> <t>anti-Myc</t> beads was performed in HEK 293T cells co-transfected with wsv156-Myc and Cq TOMM70-HA. ( E ) Wsv156 expression induced mitochondrial aggregation. Confocal microscopy of HEK 293T cells showed co-localization of wsv156-GFP (green) with the mitochondrial marker Hs COX IV (red). Cells expressing GFP alone or wsv152-GFP served as controls. ( F ) Wsv156 knockdown attenuated WSSV-induced mitochondrial aggregation. Gene silencing of wsv156 in Hpt cells reduced mitochondrial clustering (red) compared to control cells at 12 hpi. NT, non-targeting dsRNA (GFP dsRNA) control; RNAi wsv156, wsv156-specific dsRNA. The right panel quantifies mitochondrial aggregation using the CI and the percentage of cells containing mito-aggresomes. Nuclei were stained with DAPI. **, P < 0.01.
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Wsv156 targets mitochondria via Cq TOMM70 and triggers mito-aggresome formation. ( A ) Wsv156 expression increased following WSSV infection. Temporal expression of wsv156 at the mRNA (left panel) and protein level (right panel) was assessed by RT-qPCR and western blot, respectively, in Hpt cells after WSSV infection. ( B ) Wsv156 localized to mitochondria. Endogenous wsv156 protein in Hpt cells (left panel) and overexpressed wsv156 in HEK 293T cells (right panel) both predominantly co-fractionated with mitochondria. ( C ) Identification of host mitochondrial proteins interacting with wsv156 by affinity purification-mass spectrometry (AP-MS). ( D ) Wsv156 interacted with Cq TOMM70. Co-IP <t>with</t> <t>anti-Myc</t> beads was performed in HEK 293T cells co-transfected with wsv156-Myc and Cq TOMM70-HA. ( E ) Wsv156 expression induced mitochondrial aggregation. Confocal microscopy of HEK 293T cells showed co-localization of wsv156-GFP (green) with the mitochondrial marker Hs COX IV (red). Cells expressing GFP alone or wsv152-GFP served as controls. ( F ) Wsv156 knockdown attenuated WSSV-induced mitochondrial aggregation. Gene silencing of wsv156 in Hpt cells reduced mitochondrial clustering (red) compared to control cells at 12 hpi. NT, non-targeting dsRNA (GFP dsRNA) control; RNAi wsv156, wsv156-specific dsRNA. The right panel quantifies mitochondrial aggregation using the CI and the percentage of cells containing mito-aggresomes. Nuclei were stained with DAPI. **, P < 0.01.
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Wsv156 targets mitochondria via Cq TOMM70 and triggers mito-aggresome formation. ( A ) Wsv156 expression increased following WSSV infection. Temporal expression of wsv156 at the mRNA (left panel) and protein level (right panel) was assessed by RT-qPCR and western blot, respectively, in Hpt cells after WSSV infection. ( B ) Wsv156 localized to mitochondria. Endogenous wsv156 protein in Hpt cells (left panel) and overexpressed wsv156 in HEK 293T cells (right panel) both predominantly co-fractionated with mitochondria. ( C ) Identification of host mitochondrial proteins interacting with wsv156 by affinity purification-mass spectrometry (AP-MS). ( D ) Wsv156 interacted with Cq TOMM70. Co-IP <t>with</t> <t>anti-Myc</t> beads was performed in HEK 293T cells co-transfected with wsv156-Myc and Cq TOMM70-HA. ( E ) Wsv156 expression induced mitochondrial aggregation. Confocal microscopy of HEK 293T cells showed co-localization of wsv156-GFP (green) with the mitochondrial marker Hs COX IV (red). Cells expressing GFP alone or wsv152-GFP served as controls. ( F ) Wsv156 knockdown attenuated WSSV-induced mitochondrial aggregation. Gene silencing of wsv156 in Hpt cells reduced mitochondrial clustering (red) compared to control cells at 12 hpi. NT, non-targeting dsRNA (GFP dsRNA) control; RNAi wsv156, wsv156-specific dsRNA. The right panel quantifies mitochondrial aggregation using the CI and the percentage of cells containing mito-aggresomes. Nuclei were stained with DAPI. **, P < 0.01.
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Wsv156 targets mitochondria via Cq TOMM70 and triggers mito-aggresome formation. ( A ) Wsv156 expression increased following WSSV infection. Temporal expression of wsv156 at the mRNA (left panel) and protein level (right panel) was assessed by RT-qPCR and western blot, respectively, in Hpt cells after WSSV infection. ( B ) Wsv156 localized to mitochondria. Endogenous wsv156 protein in Hpt cells (left panel) and overexpressed wsv156 in HEK 293T cells (right panel) both predominantly co-fractionated with mitochondria. ( C ) Identification of host mitochondrial proteins interacting with wsv156 by affinity purification-mass spectrometry (AP-MS). ( D ) Wsv156 interacted with Cq TOMM70. Co-IP <t>with</t> <t>anti-Myc</t> beads was performed in HEK 293T cells co-transfected with wsv156-Myc and Cq TOMM70-HA. ( E ) Wsv156 expression induced mitochondrial aggregation. Confocal microscopy of HEK 293T cells showed co-localization of wsv156-GFP (green) with the mitochondrial marker Hs COX IV (red). Cells expressing GFP alone or wsv152-GFP served as controls. ( F ) Wsv156 knockdown attenuated WSSV-induced mitochondrial aggregation. Gene silencing of wsv156 in Hpt cells reduced mitochondrial clustering (red) compared to control cells at 12 hpi. NT, non-targeting dsRNA (GFP dsRNA) control; RNAi wsv156, wsv156-specific dsRNA. The right panel quantifies mitochondrial aggregation using the CI and the percentage of cells containing mito-aggresomes. Nuclei were stained with DAPI. **, P < 0.01.
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Wsv156 targets mitochondria via Cq TOMM70 and triggers mito-aggresome formation. ( A ) Wsv156 expression increased following WSSV infection. Temporal expression of wsv156 at the mRNA (left panel) and protein level (right panel) was assessed by RT-qPCR and western blot, respectively, in Hpt cells after WSSV infection. ( B ) Wsv156 localized to mitochondria. Endogenous wsv156 protein in Hpt cells (left panel) and overexpressed wsv156 in HEK 293T cells (right panel) both predominantly co-fractionated with mitochondria. ( C ) Identification of host mitochondrial proteins interacting with wsv156 by affinity purification-mass spectrometry (AP-MS). ( D ) Wsv156 interacted with Cq TOMM70. Co-IP <t>with</t> <t>anti-Myc</t> beads was performed in HEK 293T cells co-transfected with wsv156-Myc and Cq TOMM70-HA. ( E ) Wsv156 expression induced mitochondrial aggregation. Confocal microscopy of HEK 293T cells showed co-localization of wsv156-GFP (green) with the mitochondrial marker Hs COX IV (red). Cells expressing GFP alone or wsv152-GFP served as controls. ( F ) Wsv156 knockdown attenuated WSSV-induced mitochondrial aggregation. Gene silencing of wsv156 in Hpt cells reduced mitochondrial clustering (red) compared to control cells at 12 hpi. NT, non-targeting dsRNA (GFP dsRNA) control; RNAi wsv156, wsv156-specific dsRNA. The right panel quantifies mitochondrial aggregation using the CI and the percentage of cells containing mito-aggresomes. Nuclei were stained with DAPI. **, P < 0.01.
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Wsv156 targets mitochondria via Cq TOMM70 and triggers mito-aggresome formation. ( A ) Wsv156 expression increased following WSSV infection. Temporal expression of wsv156 at the mRNA (left panel) and protein level (right panel) was assessed by RT-qPCR and western blot, respectively, in Hpt cells after WSSV infection. ( B ) Wsv156 localized to mitochondria. Endogenous wsv156 protein in Hpt cells (left panel) and overexpressed wsv156 in HEK 293T cells (right panel) both predominantly co-fractionated with mitochondria. ( C ) Identification of host mitochondrial proteins interacting with wsv156 by affinity purification-mass spectrometry (AP-MS). ( D ) Wsv156 interacted with Cq TOMM70. Co-IP <t>with</t> <t>anti-Myc</t> beads was performed in HEK 293T cells co-transfected with wsv156-Myc and Cq TOMM70-HA. ( E ) Wsv156 expression induced mitochondrial aggregation. Confocal microscopy of HEK 293T cells showed co-localization of wsv156-GFP (green) with the mitochondrial marker Hs COX IV (red). Cells expressing GFP alone or wsv152-GFP served as controls. ( F ) Wsv156 knockdown attenuated WSSV-induced mitochondrial aggregation. Gene silencing of wsv156 in Hpt cells reduced mitochondrial clustering (red) compared to control cells at 12 hpi. NT, non-targeting dsRNA (GFP dsRNA) control; RNAi wsv156, wsv156-specific dsRNA. The right panel quantifies mitochondrial aggregation using the CI and the percentage of cells containing mito-aggresomes. Nuclei were stained with DAPI. **, P < 0.01.
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Wsv156 targets mitochondria via Cq TOMM70 and triggers mito-aggresome formation. ( A ) Wsv156 expression increased following WSSV infection. Temporal expression of wsv156 at the mRNA (left panel) and protein level (right panel) was assessed by RT-qPCR and western blot, respectively, in Hpt cells after WSSV infection. ( B ) Wsv156 localized to mitochondria. Endogenous wsv156 protein in Hpt cells (left panel) and overexpressed wsv156 in HEK 293T cells (right panel) both predominantly co-fractionated with mitochondria. ( C ) Identification of host mitochondrial proteins interacting with wsv156 by affinity purification-mass spectrometry (AP-MS). ( D ) Wsv156 interacted with Cq TOMM70. Co-IP <t>with</t> <t>anti-Myc</t> beads was performed in HEK 293T cells co-transfected with wsv156-Myc and Cq TOMM70-HA. ( E ) Wsv156 expression induced mitochondrial aggregation. Confocal microscopy of HEK 293T cells showed co-localization of wsv156-GFP (green) with the mitochondrial marker Hs COX IV (red). Cells expressing GFP alone or wsv152-GFP served as controls. ( F ) Wsv156 knockdown attenuated WSSV-induced mitochondrial aggregation. Gene silencing of wsv156 in Hpt cells reduced mitochondrial clustering (red) compared to control cells at 12 hpi. NT, non-targeting dsRNA (GFP dsRNA) control; RNAi wsv156, wsv156-specific dsRNA. The right panel quantifies mitochondrial aggregation using the CI and the percentage of cells containing mito-aggresomes. Nuclei were stained with DAPI. **, P < 0.01.
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Image Search Results


Wsv156 targets mitochondria via Cq TOMM70 and triggers mito-aggresome formation. ( A ) Wsv156 expression increased following WSSV infection. Temporal expression of wsv156 at the mRNA (left panel) and protein level (right panel) was assessed by RT-qPCR and western blot, respectively, in Hpt cells after WSSV infection. ( B ) Wsv156 localized to mitochondria. Endogenous wsv156 protein in Hpt cells (left panel) and overexpressed wsv156 in HEK 293T cells (right panel) both predominantly co-fractionated with mitochondria. ( C ) Identification of host mitochondrial proteins interacting with wsv156 by affinity purification-mass spectrometry (AP-MS). ( D ) Wsv156 interacted with Cq TOMM70. Co-IP with anti-Myc beads was performed in HEK 293T cells co-transfected with wsv156-Myc and Cq TOMM70-HA. ( E ) Wsv156 expression induced mitochondrial aggregation. Confocal microscopy of HEK 293T cells showed co-localization of wsv156-GFP (green) with the mitochondrial marker Hs COX IV (red). Cells expressing GFP alone or wsv152-GFP served as controls. ( F ) Wsv156 knockdown attenuated WSSV-induced mitochondrial aggregation. Gene silencing of wsv156 in Hpt cells reduced mitochondrial clustering (red) compared to control cells at 12 hpi. NT, non-targeting dsRNA (GFP dsRNA) control; RNAi wsv156, wsv156-specific dsRNA. The right panel quantifies mitochondrial aggregation using the CI and the percentage of cells containing mito-aggresomes. Nuclei were stained with DAPI. **, P < 0.01.

Journal: Journal of Virology

Article Title: The white spot syndrome virus wsv156 protein hijacks Parkin-dependent mitophagy to promote viral infection

doi: 10.1128/jvi.00418-26

Figure Lengend Snippet: Wsv156 targets mitochondria via Cq TOMM70 and triggers mito-aggresome formation. ( A ) Wsv156 expression increased following WSSV infection. Temporal expression of wsv156 at the mRNA (left panel) and protein level (right panel) was assessed by RT-qPCR and western blot, respectively, in Hpt cells after WSSV infection. ( B ) Wsv156 localized to mitochondria. Endogenous wsv156 protein in Hpt cells (left panel) and overexpressed wsv156 in HEK 293T cells (right panel) both predominantly co-fractionated with mitochondria. ( C ) Identification of host mitochondrial proteins interacting with wsv156 by affinity purification-mass spectrometry (AP-MS). ( D ) Wsv156 interacted with Cq TOMM70. Co-IP with anti-Myc beads was performed in HEK 293T cells co-transfected with wsv156-Myc and Cq TOMM70-HA. ( E ) Wsv156 expression induced mitochondrial aggregation. Confocal microscopy of HEK 293T cells showed co-localization of wsv156-GFP (green) with the mitochondrial marker Hs COX IV (red). Cells expressing GFP alone or wsv152-GFP served as controls. ( F ) Wsv156 knockdown attenuated WSSV-induced mitochondrial aggregation. Gene silencing of wsv156 in Hpt cells reduced mitochondrial clustering (red) compared to control cells at 12 hpi. NT, non-targeting dsRNA (GFP dsRNA) control; RNAi wsv156, wsv156-specific dsRNA. The right panel quantifies mitochondrial aggregation using the CI and the percentage of cells containing mito-aggresomes. Nuclei were stained with DAPI. **, P < 0.01.

Article Snippet: Co-IP was performed using anti-Myc magnetic beads (MedChemExpress, HY-K0206) as previously described ( ).

Techniques: Expressing, Infection, Quantitative RT-PCR, Western Blot, Affinity Purification, Mass Spectrometry, Protein-Protein interactions, Co-Immunoprecipitation Assay, Transfection, Confocal Microscopy, Marker, Knockdown, Control, Staining