Shrimp pmo-miR-750 regulates the expression of sarcoplasmic calcium-binding protein facilitating virus infection in Penaeus monodon
MicroRNAs (miRNAs) regulate gene expression post-transcriptionally and play crucial roles in antiviral responses. Penaeus monodon miR-750 (pmo-miR-750) was found to be strongly up-regulated in the late phase of white spot syndrome virus (WSSV) infection, but its function remains uncharacterized. Her...
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Veröffentlicht in: | Fish & shellfish immunology 2022-10, Vol.129, p.74-84 |
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description | MicroRNAs (miRNAs) regulate gene expression post-transcriptionally and play crucial roles in antiviral responses. Penaeus monodon miR-750 (pmo-miR-750) was found to be strongly up-regulated in the late phase of white spot syndrome virus (WSSV) infection, but its function remains uncharacterized. Herein, the targets that were translationally down-regulated in the shrimp stomach following a pmo-miR-750 mimic injection were identified using two-dimensional gel electrophoresis. Sarcoplasmic calcium-binding protein (Scp) and actin1 (Act1) were revealed to be down-regulated protein spots. The genuine binding of pmo-miR-750 mimic to Scp but not Act1 mRNA was validated in vitro. In addition, a negative correlation between the Scp transcript and pmo-miR-750 expression level in WSSV-infected P. monodon stomach implies that pmo-miR-750 regulates Scp expression in vivo. When injected into WSSV-infected shrimp, the pmo-miR-750 mimic suppressed Scp expression but significantly increased the WSSV copy number. Consistent with the miRNA mimic-mediated Scp suppression, the loss of function assay of Scp in WSSV-challenged shrimp by RNA interference revealed a decreased survival rate with a dramatic increase in viral copy number. Besides that, apoptosis was activated in the hemocytes of the Scp knockdown shrimp upon WSSV infection. Collectively, our findings reveal that up-regulated pmo-miR-750 suppresses Scp expression at both the transcript and protein levels in the late stage of WSSV infection, which contributes to modulating apoptosis and eventually enabling viral propagation.
•The pmo-miR-750 mimic introduced into shrimp reduced Scp and Act1 expression.•Scp participated in regulating apoptosis in shrimp upon WSSV infection.•The upregulation of pmo-miR-750 upon WSSV infection suppressed Scp expression.•Apoptosis induced by the pmo-miR-750/Scp interaction enhances WSSV replication. |
doi_str_mv | 10.1016/j.fsi.2022.08.046 |
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•The pmo-miR-750 mimic introduced into shrimp reduced Scp and Act1 expression.•Scp participated in regulating apoptosis in shrimp upon WSSV infection.•The upregulation of pmo-miR-750 upon WSSV infection suppressed Scp expression.•Apoptosis induced by the pmo-miR-750/Scp interaction enhances WSSV replication.</description><identifier>ISSN: 1050-4648</identifier><identifier>EISSN: 1095-9947</identifier><identifier>DOI: 10.1016/j.fsi.2022.08.046</identifier><language>eng</language><publisher>Elsevier Ltd</publisher><subject>Penaeus monodon ; pmo-miR-750 ; Viral-responsive miRNA ; White spot syndrome virus</subject><ispartof>Fish & shellfish immunology, 2022-10, Vol.129, p.74-84</ispartof><rights>2022 Elsevier Ltd</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c260t-c66be649ce3cdc067f9fe666a73c034d72dc5bee86bc54fe118858146c23f9153</citedby><cites>FETCH-LOGICAL-c260t-c66be649ce3cdc067f9fe666a73c034d72dc5bee86bc54fe118858146c23f9153</cites><orcidid>0000-0002-0241-8238</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.fsi.2022.08.046$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids></links><search><creatorcontrib>Kanoksinwuttipong, Nichaphat</creatorcontrib><creatorcontrib>Jaree, Phattarunda</creatorcontrib><creatorcontrib>Somboonwiwat, Kunlaya</creatorcontrib><title>Shrimp pmo-miR-750 regulates the expression of sarcoplasmic calcium-binding protein facilitating virus infection in Penaeus monodon</title><title>Fish & shellfish immunology</title><description>MicroRNAs (miRNAs) regulate gene expression post-transcriptionally and play crucial roles in antiviral responses. Penaeus monodon miR-750 (pmo-miR-750) was found to be strongly up-regulated in the late phase of white spot syndrome virus (WSSV) infection, but its function remains uncharacterized. Herein, the targets that were translationally down-regulated in the shrimp stomach following a pmo-miR-750 mimic injection were identified using two-dimensional gel electrophoresis. Sarcoplasmic calcium-binding protein (Scp) and actin1 (Act1) were revealed to be down-regulated protein spots. The genuine binding of pmo-miR-750 mimic to Scp but not Act1 mRNA was validated in vitro. In addition, a negative correlation between the Scp transcript and pmo-miR-750 expression level in WSSV-infected P. monodon stomach implies that pmo-miR-750 regulates Scp expression in vivo. When injected into WSSV-infected shrimp, the pmo-miR-750 mimic suppressed Scp expression but significantly increased the WSSV copy number. Consistent with the miRNA mimic-mediated Scp suppression, the loss of function assay of Scp in WSSV-challenged shrimp by RNA interference revealed a decreased survival rate with a dramatic increase in viral copy number. Besides that, apoptosis was activated in the hemocytes of the Scp knockdown shrimp upon WSSV infection. Collectively, our findings reveal that up-regulated pmo-miR-750 suppresses Scp expression at both the transcript and protein levels in the late stage of WSSV infection, which contributes to modulating apoptosis and eventually enabling viral propagation.
•The pmo-miR-750 mimic introduced into shrimp reduced Scp and Act1 expression.•Scp participated in regulating apoptosis in shrimp upon WSSV infection.•The upregulation of pmo-miR-750 upon WSSV infection suppressed Scp expression.•Apoptosis induced by the pmo-miR-750/Scp interaction enhances WSSV replication.</description><subject>Penaeus monodon</subject><subject>pmo-miR-750</subject><subject>Viral-responsive miRNA</subject><subject>White spot syndrome virus</subject><issn>1050-4648</issn><issn>1095-9947</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><recordid>eNp9kM1O3TAQhSNEJeilD9Cdl2ySjvPjJGKFUAuVkFqVsrZ8J2OYq8RO7QTRdV8cR7drVjM6M-do5suyzxIKCVJ9ORQ2clFCWRbQFVCrk-xcQt_kfV-3p1vfQF6rujvLPsZ4AABVKTjP_j08B55mMU8-n_hX3jYgAj2to1koiuWZBL3OgWJk74S3IpqAfh5NnBgFmhF5nfI9u4Hdk5iDX4idsAZ55MUsm_jCYY2CnSVctpA0_0nOUBIn7_zg3UX2wZox0qf_dZc9fvv6--Yuv_9x-_3m-j7HUsGSo1J7UnWPVOGAoFrbW1JKmbZCqOqhLQds9kSd2mNTW5Ky65pO1grLyvayqXbZ5TE33flnpbjoiSPSOBpHfo26bKFVsoOyTavyuIrBxxjI6jlhMuGvlqA34PqgE3C9AdfQ6QQ8ea6OHko_vDAFHZHJIQ0c0u968PyO-w1WIou9</recordid><startdate>202210</startdate><enddate>202210</enddate><creator>Kanoksinwuttipong, Nichaphat</creator><creator>Jaree, Phattarunda</creator><creator>Somboonwiwat, Kunlaya</creator><general>Elsevier Ltd</general><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-0241-8238</orcidid></search><sort><creationdate>202210</creationdate><title>Shrimp pmo-miR-750 regulates the expression of sarcoplasmic calcium-binding protein facilitating virus infection in Penaeus monodon</title><author>Kanoksinwuttipong, Nichaphat ; Jaree, Phattarunda ; Somboonwiwat, Kunlaya</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c260t-c66be649ce3cdc067f9fe666a73c034d72dc5bee86bc54fe118858146c23f9153</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>Penaeus monodon</topic><topic>pmo-miR-750</topic><topic>Viral-responsive miRNA</topic><topic>White spot syndrome virus</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kanoksinwuttipong, Nichaphat</creatorcontrib><creatorcontrib>Jaree, Phattarunda</creatorcontrib><creatorcontrib>Somboonwiwat, Kunlaya</creatorcontrib><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Fish & shellfish immunology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kanoksinwuttipong, Nichaphat</au><au>Jaree, Phattarunda</au><au>Somboonwiwat, Kunlaya</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Shrimp pmo-miR-750 regulates the expression of sarcoplasmic calcium-binding protein facilitating virus infection in Penaeus monodon</atitle><jtitle>Fish & shellfish immunology</jtitle><date>2022-10</date><risdate>2022</risdate><volume>129</volume><spage>74</spage><epage>84</epage><pages>74-84</pages><issn>1050-4648</issn><eissn>1095-9947</eissn><abstract>MicroRNAs (miRNAs) regulate gene expression post-transcriptionally and play crucial roles in antiviral responses. Penaeus monodon miR-750 (pmo-miR-750) was found to be strongly up-regulated in the late phase of white spot syndrome virus (WSSV) infection, but its function remains uncharacterized. Herein, the targets that were translationally down-regulated in the shrimp stomach following a pmo-miR-750 mimic injection were identified using two-dimensional gel electrophoresis. Sarcoplasmic calcium-binding protein (Scp) and actin1 (Act1) were revealed to be down-regulated protein spots. The genuine binding of pmo-miR-750 mimic to Scp but not Act1 mRNA was validated in vitro. In addition, a negative correlation between the Scp transcript and pmo-miR-750 expression level in WSSV-infected P. monodon stomach implies that pmo-miR-750 regulates Scp expression in vivo. When injected into WSSV-infected shrimp, the pmo-miR-750 mimic suppressed Scp expression but significantly increased the WSSV copy number. Consistent with the miRNA mimic-mediated Scp suppression, the loss of function assay of Scp in WSSV-challenged shrimp by RNA interference revealed a decreased survival rate with a dramatic increase in viral copy number. Besides that, apoptosis was activated in the hemocytes of the Scp knockdown shrimp upon WSSV infection. Collectively, our findings reveal that up-regulated pmo-miR-750 suppresses Scp expression at both the transcript and protein levels in the late stage of WSSV infection, which contributes to modulating apoptosis and eventually enabling viral propagation.
•The pmo-miR-750 mimic introduced into shrimp reduced Scp and Act1 expression.•Scp participated in regulating apoptosis in shrimp upon WSSV infection.•The upregulation of pmo-miR-750 upon WSSV infection suppressed Scp expression.•Apoptosis induced by the pmo-miR-750/Scp interaction enhances WSSV replication.</abstract><pub>Elsevier Ltd</pub><doi>10.1016/j.fsi.2022.08.046</doi><tpages>11</tpages><orcidid>https://orcid.org/0000-0002-0241-8238</orcidid></addata></record> |
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subjects | Penaeus monodon pmo-miR-750 Viral-responsive miRNA White spot syndrome virus |
title | Shrimp pmo-miR-750 regulates the expression of sarcoplasmic calcium-binding protein facilitating virus infection in Penaeus monodon |
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