Transmissible Gastroenteritis Virus (TGEV) Infection Alters the Expression of Cellular MicroRNA Species That Affect Transcription of TGEV Gene 7
Identifieur interne : 000009 ( Pmc/Curation ); précédent : 000008; suivant : 000010Transmissible Gastroenteritis Virus (TGEV) Infection Alters the Expression of Cellular MicroRNA Species That Affect Transcription of TGEV Gene 7
Auteurs : Xiangjun Song ; Xiaomin Zhao ; Yong Huang ; Hailing Xiang ; Wenlong Zhang ; Dewen TongSource :
- International Journal of Biological Sciences [ 1449-2288 ] ; 2015.
Abstract
Transmissible gastroenteritis virus (TGEV) is a member of
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DOI: 10.7150/ijbs.11585
PubMed: 26157346
PubMed Central: 4495409
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Transmissible Gastroenteritis Virus (TGEV) Infection Alters the Expression of Cellular MicroRNA Species That Affect Transcription of TGEV Gene 7</title><author><name sortKey="Song, Xiangjun" sort="Song, Xiangjun" uniqKey="Song X" first="Xiangjun" last="Song">Xiangjun Song</name></author><author><name sortKey="Zhao, Xiaomin" sort="Zhao, Xiaomin" uniqKey="Zhao X" first="Xiaomin" last="Zhao">Xiaomin Zhao</name></author><author><name sortKey="Huang, Yong" sort="Huang, Yong" uniqKey="Huang Y" first="Yong" last="Huang">Yong Huang</name></author><author><name sortKey="Xiang, Hailing" sort="Xiang, Hailing" uniqKey="Xiang H" first="Hailing" last="Xiang">Hailing Xiang</name></author><author><name sortKey="Zhang, Wenlong" sort="Zhang, Wenlong" uniqKey="Zhang W" first="Wenlong" last="Zhang">Wenlong Zhang</name></author><author><name sortKey="Tong, Dewen" sort="Tong, Dewen" uniqKey="Tong D" first="Dewen" last="Tong">Dewen Tong</name></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">26157346</idno><idno type="pmc">4495409</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4495409</idno><idno type="RBID">PMC:4495409</idno><idno type="doi">10.7150/ijbs.11585</idno><date when="2015">2015</date><idno type="wicri:Area/Pmc/Corpus">000009</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000009</idno><idno type="wicri:Area/Pmc/Curation">000009</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000009</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Transmissible Gastroenteritis Virus (TGEV) Infection Alters the Expression of Cellular MicroRNA Species That Affect Transcription of TGEV Gene 7</title><author><name sortKey="Song, Xiangjun" sort="Song, Xiangjun" uniqKey="Song X" first="Xiangjun" last="Song">Xiangjun Song</name></author><author><name sortKey="Zhao, Xiaomin" sort="Zhao, Xiaomin" uniqKey="Zhao X" first="Xiaomin" last="Zhao">Xiaomin Zhao</name></author><author><name sortKey="Huang, Yong" sort="Huang, Yong" uniqKey="Huang Y" first="Yong" last="Huang">Yong Huang</name></author><author><name sortKey="Xiang, Hailing" sort="Xiang, Hailing" uniqKey="Xiang H" first="Hailing" last="Xiang">Hailing Xiang</name></author><author><name sortKey="Zhang, Wenlong" sort="Zhang, Wenlong" uniqKey="Zhang W" first="Wenlong" last="Zhang">Wenlong Zhang</name></author><author><name sortKey="Tong, Dewen" sort="Tong, Dewen" uniqKey="Tong D" first="Dewen" last="Tong">Dewen Tong</name></author></analytic><series><title level="j">International Journal of Biological Sciences</title><idno type="eISSN">1449-2288</idno><imprint><date when="2015">2015</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Transmissible gastroenteritis virus (TGEV) is a member of <italic>Coronaviridae </italic>family. TGEV infection has emerged as a major cause of severe gastroenteritis and leads to alterations of many cellular processes. Meanwhile, the pathogenic mechanism of TGEV is still unclear. microRNAs (miRNAs) are a novel class of small non-coding RNAs which are involved in the regulation of numerous biological processes such as viral infection and cell apoptosis. Accumulating data show that miRNAs are involved in the process of coronavirus infection such as replication of severe acute respiratory syndrome coronavirus (SARS-CoV). However, the link between miRNAs and TGEV infection is unknown. In this study, we performed microRNA microarray assay and predicted targets of altered miRNAs. The results showed TGEV infection caused the change of miRNAs profile. Then we selected miR-4331 for further analysis and subsequently identified cell division cycle-associated protein 7 (CDCA7) as the target of miR-4331. Moreover, miR-4331 showed the ability to inhibit transcription of TGEV gene 7 (a non-structure gene) via directly targeting CDCA7. In conclusion, differentially expressed miR-4331 that is caused by TGEV infection can suppress transcription of TGEV gene 7 via targeting cellular CDCA7. Our key finding is that TGEV selectively manipulates the expression of some cellular miRNAs to regulate its subgenomic transcription.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Wilson, Rc" uniqKey="Wilson R">RC Wilson</name></author><author><name sortKey="Doudna, Ja" uniqKey="Doudna J">JA Doudna</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Sun, W" uniqKey="Sun W">W Sun</name></author><author><name sortKey="Julie Li, Ys" uniqKey="Julie Li Y">YS Julie Li</name></author><author><name sortKey="Huang, Hd" uniqKey="Huang H">HD Huang</name></author><author><name sortKey="Shyy, Jy" uniqKey="Shyy J">JY Shyy</name></author><author><name sortKey="Chien, S" uniqKey="Chien S">S Chien</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Valinezhad Orang, A" uniqKey="Valinezhad Orang A">A Valinezhad Orang</name></author><author><name sortKey="Safaralizadeh, R" 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Int J Biol Sci</journal-id><journal-id journal-id-type="iso-abbrev">Int. J. Biol. Sci</journal-id><journal-id journal-id-type="publisher-id">ijbs</journal-id><journal-title-group><journal-title>International Journal of Biological Sciences</journal-title></journal-title-group><issn pub-type="epub">1449-2288</issn><publisher><publisher-name>Ivyspring International Publisher</publisher-name><publisher-loc>Sydney</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">26157346</article-id><article-id pub-id-type="pmc">4495409</article-id><article-id pub-id-type="doi">10.7150/ijbs.11585</article-id><article-id pub-id-type="publisher-id">ijbsv11p0913</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Paper</subject></subj-group></article-categories><title-group><article-title>Transmissible Gastroenteritis Virus (TGEV) Infection Alters the Expression of Cellular MicroRNA Species That Affect Transcription of TGEV Gene 7</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Song</surname><given-names>Xiangjun</given-names></name><xref ref-type="author-notes" rid="FNA_star">*</xref></contrib><contrib contrib-type="author"><name><surname>Zhao</surname><given-names>Xiaomin</given-names></name><xref ref-type="author-notes" rid="FNA_star">*</xref></contrib><contrib contrib-type="author"><name><surname>Huang</surname><given-names>Yong</given-names></name></contrib><contrib contrib-type="author"><name><surname>Xiang</surname><given-names>Hailing</given-names></name></contrib><contrib contrib-type="author"><name><surname>Zhang</surname><given-names>Wenlong</given-names></name></contrib><contrib contrib-type="author"><name><surname>Tong</surname><given-names>Dewen</given-names></name><xref ref-type="corresp" rid="FNA_envelop">✉</xref></contrib></contrib-group><aff>College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, P.R. China</aff><author-notes><corresp id="FNA_envelop">✉ Corresponding author: Prof. Dewen Tong. Telephone: +86-29-87091622; Fax: +86-29-87091032; E-mail: <email>dwtong@nwsuaf.edu.cn</email></corresp><fn fn-type="equal" id="FNA_star"><p>* These authors contributed equally to this work.</p></fn><fn fn-type="conflict"><p>Competing interests: The authors have declared that no competing interest exists.</p></fn></author-notes><pub-date pub-type="collection"><year>2015</year></pub-date><pub-date pub-type="epub"><day>7</day><month>6</month><year>2015</year></pub-date><volume>11</volume><issue>8</issue><fpage>913</fpage><lpage>922</lpage><history><date date-type="received"><day>13</day><month>1</month><year>2015</year></date><date date-type="accepted"><day>14</day><month>5</month><year>2015</year></date></history><permissions><copyright-statement>© 2015 Ivyspring International Publisher. Reproduction is permitted for personal, noncommercial use, provided that the article is in whole, unmodified, and properly cited. See http://ivyspring.com/terms for terms and conditions.</copyright-statement><copyright-year>2015</copyright-year></permissions><abstract><p>Transmissible gastroenteritis virus (TGEV) is a member of <italic>Coronaviridae </italic>family. TGEV infection has emerged as a major cause of severe gastroenteritis and leads to alterations of many cellular processes. Meanwhile, the pathogenic mechanism of TGEV is still unclear. microRNAs (miRNAs) are a novel class of small non-coding RNAs which are involved in the regulation of numerous biological processes such as viral infection and cell apoptosis. Accumulating data show that miRNAs are involved in the process of coronavirus infection such as replication of severe acute respiratory syndrome coronavirus (SARS-CoV). However, the link between miRNAs and TGEV infection is unknown. In this study, we performed microRNA microarray assay and predicted targets of altered miRNAs. The results showed TGEV infection caused the change of miRNAs profile. Then we selected miR-4331 for further analysis and subsequently identified cell division cycle-associated protein 7 (CDCA7) as the target of miR-4331. Moreover, miR-4331 showed the ability to inhibit transcription of TGEV gene 7 (a non-structure gene) via directly targeting CDCA7. In conclusion, differentially expressed miR-4331 that is caused by TGEV infection can suppress transcription of TGEV gene 7 via targeting cellular CDCA7. Our key finding is that TGEV selectively manipulates the expression of some cellular miRNAs to regulate its subgenomic transcription.</p></abstract><kwd-group><kwd>TGEV</kwd><kwd>microRNAs (miRNAs)</kwd><kwd>TGEV</kwd><kwd>CDCA7</kwd><kwd>Gene 7</kwd><kwd>viral transcription</kwd></kwd-group></article-meta></front><floats-group><fig id="F1" position="float"><label>Fig 1</label><caption><p>miRNAs expression profile in TGEV-infected PK-15 cells. (A) Heat map analysis showing differentially expressed miRNAs in PK-15 cells infected with TGEV at 24 h. Red indicates higher expression and green indicates lower expression. Each colored block represents the expression of one miRNA (labeled on the right) in the indicated sample. The map showed all significantly differential expressed miRNAs in three independent samples (p < 0.01). (B) Verification of miRNAs microarray by real-time PCR. The fold change was determined using the 2<sup>-ΔΔCt</sup> method and all miRNAs expression values were normalized to endogenous U6. Data from real-time PCR were shown as mean ±standard deviation (S.D.) of three independent experiments. Similar results were obtained in three independent experiments.</p></caption><graphic xlink:href="ijbsv11p0913g001"></graphic></fig><fig id="F2" position="float"><label>Fig 2</label><caption><p>The effect of miR-4331 on transcription of TGEV gene 7. (A) miR-4331 levels in PK-15 cells transfected with mimics or inhibitors and infected with TGEV at 12 and 24 hpi. The expression of miR-4331 was measured by real-time PCR (Normalized to U6 and in reference to the level of the control). (B) The effect of miR-4331 on transcription of TGEV gene 7 at 12 and 24 hpi. The effect of miR-4331 on TGEV transcription of gene 7 was assessed by real-time PCR analysis (Normalized to gRNA and in reference to the level of the control). Data represent mean ±S.D. of three independent experiments. * P < 0.05 in comparison with the control. ** P < 0.01 in comparison with the control.</p></caption><graphic xlink:href="ijbsv11p0913g002"></graphic></fig><fig id="F3" position="float"><label>Fig 3</label><caption><p>Identification of miR-4331 target. (A) The effect of miR-4331 on 3' UTRs of 16 potential target genes measured by dual-luciferase reporter assay (normalized to firefly). (B) Bioinformatic prediction of interaction between miR-4331 and the 3' UTRs of swine CDCA7. For each schematic, the upper sequence is the binding site of miR-4331 in 3ʹUTRs of swine CDCA7, the middle is the sequence of mature miR-4331, and the lower sequence is the mutated sequence of CDCA7 3ʹ UTR. The seed sequence is underlined. (C) Schematic overview of the putative binding sites or mutations of miR-4331 binding-sites in 3' UTR of CDCA7 mRNA. The locations of the potential binding sites or their mutations are presented by blank boxes. (D) The CDCA7 luciferase reporter construct was co-transfected with miR-4331 mimics (or negative control) or miR-4331 inhibitors (or negative control) into PK-15 cells (normalized to the firefly luciferase activity). Data are expressed as relative luciferase activities to control. (E) The specific binding activity of miR-4331 mimics and inhibitors (normalized to the firefly luciferase activity). (F) Western blot analysis of the effect of miR-4331CDCA7 in cells transfected with miR-4331 mimics or mimics control. (G) Western blot analysis of CDCA7 in cells transfected with miR-4331 inhibitors or inhibitors control. (H) Western blot analysis of endogenous CDCA7 protein level in TGEV-infected cells at 24 hpi. (I) Western blot analysis of endogenous CDCA7 protein level in TGEV-infected cells at 24 hpi by knockdown of endogenous miR-4331 using miR-4331 inhibitors. Data represent means ±S.D. of three independent experiments. * P < 0.05 in comparison with the control. ** P < 0.01 in comparison with the control.</p></caption><graphic xlink:href="ijbsv11p0913g003"></graphic><graphic xlink:href="ijbsv11p0913g004"></graphic></fig><fig id="F4" position="float"><label>Fig 4</label><caption><p>Effect of CDCA7 on transcription of TGEV gene 7. (A) Silencing effect of CDCA7 siRNA on CDCA7 expression. PK-15 cells were transfected with CDCA7-specific siRNA or irrelevant siRNA and analyzed by western blot. (B) The effect of siCDCA7-2 on CDCA7 at mRNA level. The CDCA7 mRNA level was reduced at 24 h hpt measured by real-PCR (Normalized to β-actin). (C) The effect of siCDCA7-2 on PK-15-cell viability. The cells were incubated after transfecting with 50 nM siCDCA7-2 for 48 h. Cell viability was evaluated by CCK-8 assay. (D) The overexpression effect of CDCA7 using pCI-neo-CDCA7. PK-15 cells were transfected with pCI-neo-CDCA7 or pCI-neo vector, and the expression level of CDCA7 was assessed by western blot at 48 hpt. (E) The effect of CDCA7 on transcription of TGEV gene 7. Knockdown of CDCA7 using siRNA decreased transcription of gene 7 at 12 and 24 hpi. Overexpression of CDCA7 increased transcription of gene 7 at 12 and 24 hpi. Data represent mean ±S.D. of three independent experiments. * P < 0.05 in comparison with the control. ** P < 0.01 in comparison with the control.</p></caption><graphic xlink:href="ijbsv11p0913g005"></graphic></fig></floats-group></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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