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Transmissible gastroenteritis virus (TGEV) infection alters the expression of cellular microRNA species that affect transcription of TGEV gene 7.

Song X, Zhao X, Huang Y, Xiang H, Zhang W, Tong D - Int. J. Biol. Sci. (2015)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, P.R. China.

ABSTRACT
Transmissible gastroenteritis virus (TGEV) is a member of Coronaviridae 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.

No MeSH data available.


Related in: MedlinePlus

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.
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Figure 2: 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.

Mentions: To investigate the impact of differentially expressed miRNAs on TGEV transcription, we selected miR-4331 as a representative based on p value (p < 0.01), fold change of miRNA expression (fold change > 1.5), and mean fluorescence intensities (MFI) (MFI > 400). PK-15 cells were transfected with miR-4331 mimics, mimics control, inhibitors, or inhibitors control. Subsequently all the samples and controls were infected with TGEV. The miR-4331 level of the samples was compared with the controls and normalized to U6. The miR-4331 level increased after transfection with the miRNA mimics and decreased after transfection of the inhibitors in comparison with the control (Fig. 2A). The transcription of TGEV gene 7 was decreased by overexpression of miR-4331 and increased by inhibition of miR-4331 at 12 and 24 hpi (Fig. 2B). Except for gene 7, the replication of TGEV genome and transcription of other subgenomes were not affected by miR-4331 (data are not shown).


Transmissible gastroenteritis virus (TGEV) infection alters the expression of cellular microRNA species that affect transcription of TGEV gene 7.

Song X, Zhao X, Huang Y, Xiang H, Zhang W, Tong D - Int. J. Biol. Sci. (2015)

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.
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Related In: Results  -  Collection

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Figure 2: 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.
Mentions: To investigate the impact of differentially expressed miRNAs on TGEV transcription, we selected miR-4331 as a representative based on p value (p < 0.01), fold change of miRNA expression (fold change > 1.5), and mean fluorescence intensities (MFI) (MFI > 400). PK-15 cells were transfected with miR-4331 mimics, mimics control, inhibitors, or inhibitors control. Subsequently all the samples and controls were infected with TGEV. The miR-4331 level of the samples was compared with the controls and normalized to U6. The miR-4331 level increased after transfection with the miRNA mimics and decreased after transfection of the inhibitors in comparison with the control (Fig. 2A). The transcription of TGEV gene 7 was decreased by overexpression of miR-4331 and increased by inhibition of miR-4331 at 12 and 24 hpi (Fig. 2B). Except for gene 7, the replication of TGEV genome and transcription of other subgenomes were not affected by miR-4331 (data are not shown).

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, P.R. China.

ABSTRACT
Transmissible gastroenteritis virus (TGEV) is a member of Coronaviridae 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.

No MeSH data available.


Related in: MedlinePlus