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XRN1 stalling in the 5' UTR of Hepatitis C virus and Bovine Viral Diarrhea virus is associated with dysregulated host mRNA stability.

Moon SL, Blackinton JG, Anderson JR, Dozier MK, Dodd BJ, Keene JD, Wilusz CJ, Bradrick SS, Wilusz J - PLoS Pathog. (2015)

Bottom Line: We demonstrate that both Hepatitis C virus (HCV) and Bovine Viral Diarrhea virus (BVDV) contain regions in their 5' UTRs that stall and repress the enzymatic activity of the cellular 5'-3' exoribonuclease XRN1, resulting in dramatic changes in the stability of cellular mRNAs.In the context of HCV infection, we observed globally increased stability of mRNAs resulting in significant increases in abundance of normally short-lived mRNAs encoding a variety of relevant oncogenes and angiogenesis factors.These findings suggest that non-coding regions from multiple genera of the Flaviviridae interfere with XRN1 and impact post-transcriptional processes, causing global dysregulation of cellular gene expression which may promote cell growth and pathogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado, United States of America.

ABSTRACT
We demonstrate that both Hepatitis C virus (HCV) and Bovine Viral Diarrhea virus (BVDV) contain regions in their 5' UTRs that stall and repress the enzymatic activity of the cellular 5'-3' exoribonuclease XRN1, resulting in dramatic changes in the stability of cellular mRNAs. We used biochemical assays, virus infections, and transfection of the HCV and BVDV 5' untranslated regions in the absence of other viral gene products to directly demonstrate the existence and mechanism of this novel host-virus interaction. In the context of HCV infection, we observed globally increased stability of mRNAs resulting in significant increases in abundance of normally short-lived mRNAs encoding a variety of relevant oncogenes and angiogenesis factors. These findings suggest that non-coding regions from multiple genera of the Flaviviridae interfere with XRN1 and impact post-transcriptional processes, causing global dysregulation of cellular gene expression which may promote cell growth and pathogenesis.

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Changes in gene expression also indicate XRN1 suppression and increased mRNA stability during BVDV infection.Panel A. MDBK cells were either mock infected or infected with BVDV. Total RNA samples were assayed for the indicated cellular mRNAs by qRT-PCR using ACTB as a reference transcript. The average relative abundance of FOS and JUN mRNAs from three independent infections +/− standard deviation is depicted. Panel B. Western blot analysis of total protein samples using antibodies against FOS, JUN, or GAPDH. The average quantification of the blot (relative to GAPDH) +/− standard deviation from three infections is shown. Significance was determined by t-test with * p≤0.01; **p≤0.001. Panel C. mRNA half-lives were determined using actinomycin D for the FOS and JUN mRNAs in mock-infected or BVDV-infected MDBK cells. A representative mRNA decay curve is depicted with the average +/− standard deviation of each half-life from three independent infections shown in the graphical insets. Statistical analysis was performed using a t-test with * p ≤ 0.05 and ***p≤0.005.
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ppat.1004708.g007: Changes in gene expression also indicate XRN1 suppression and increased mRNA stability during BVDV infection.Panel A. MDBK cells were either mock infected or infected with BVDV. Total RNA samples were assayed for the indicated cellular mRNAs by qRT-PCR using ACTB as a reference transcript. The average relative abundance of FOS and JUN mRNAs from three independent infections +/− standard deviation is depicted. Panel B. Western blot analysis of total protein samples using antibodies against FOS, JUN, or GAPDH. The average quantification of the blot (relative to GAPDH) +/− standard deviation from three infections is shown. Significance was determined by t-test with * p≤0.01; **p≤0.001. Panel C. mRNA half-lives were determined using actinomycin D for the FOS and JUN mRNAs in mock-infected or BVDV-infected MDBK cells. A representative mRNA decay curve is depicted with the average +/− standard deviation of each half-life from three independent infections shown in the graphical insets. Statistical analysis was performed using a t-test with * p ≤ 0.05 and ***p≤0.005.

Mentions: Finally, to generalize our observations regarding dysregulation of cellular mRNA stability in an HCV infection to another non-arthropod-borne member of the Flaviviridae, we assessed the abundance and stability of select cellular mRNAs in BVDV infected MDBK cells. As shown in Fig. 7A, the abundance of both FOS and JUN mRNAs was significantly upregulated (three to six fold) in a BVDV infection. Furthermore, the proteins encoded by these mRNAs were also upregulated in a BVDV infection (Fig. 7B) (and a similar increase in protein expression was seen in HCV infection (S4 Fig). As seen in Fig. 7C, the increase in abundance of the FOS and JUN mRNAs can largely be accounted for by the approximately five and three-fold significant increase in mRNA half-life in a BVDV infection, respectively. Importantly, XRN1 levels are not significantly changed during BVDV infection (S1B Fig). Thus, we conclude that dysregulation of cellular mRNA stability is a common occurrence in hepacivirus and pestivirus infections and may play a heretofore unforeseen role in the success of viral replication as well as viral-induced pathogenesis.


XRN1 stalling in the 5' UTR of Hepatitis C virus and Bovine Viral Diarrhea virus is associated with dysregulated host mRNA stability.

Moon SL, Blackinton JG, Anderson JR, Dozier MK, Dodd BJ, Keene JD, Wilusz CJ, Bradrick SS, Wilusz J - PLoS Pathog. (2015)

Changes in gene expression also indicate XRN1 suppression and increased mRNA stability during BVDV infection.Panel A. MDBK cells were either mock infected or infected with BVDV. Total RNA samples were assayed for the indicated cellular mRNAs by qRT-PCR using ACTB as a reference transcript. The average relative abundance of FOS and JUN mRNAs from three independent infections +/− standard deviation is depicted. Panel B. Western blot analysis of total protein samples using antibodies against FOS, JUN, or GAPDH. The average quantification of the blot (relative to GAPDH) +/− standard deviation from three infections is shown. Significance was determined by t-test with * p≤0.01; **p≤0.001. Panel C. mRNA half-lives were determined using actinomycin D for the FOS and JUN mRNAs in mock-infected or BVDV-infected MDBK cells. A representative mRNA decay curve is depicted with the average +/− standard deviation of each half-life from three independent infections shown in the graphical insets. Statistical analysis was performed using a t-test with * p ≤ 0.05 and ***p≤0.005.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4352041&req=5

ppat.1004708.g007: Changes in gene expression also indicate XRN1 suppression and increased mRNA stability during BVDV infection.Panel A. MDBK cells were either mock infected or infected with BVDV. Total RNA samples were assayed for the indicated cellular mRNAs by qRT-PCR using ACTB as a reference transcript. The average relative abundance of FOS and JUN mRNAs from three independent infections +/− standard deviation is depicted. Panel B. Western blot analysis of total protein samples using antibodies against FOS, JUN, or GAPDH. The average quantification of the blot (relative to GAPDH) +/− standard deviation from three infections is shown. Significance was determined by t-test with * p≤0.01; **p≤0.001. Panel C. mRNA half-lives were determined using actinomycin D for the FOS and JUN mRNAs in mock-infected or BVDV-infected MDBK cells. A representative mRNA decay curve is depicted with the average +/− standard deviation of each half-life from three independent infections shown in the graphical insets. Statistical analysis was performed using a t-test with * p ≤ 0.05 and ***p≤0.005.
Mentions: Finally, to generalize our observations regarding dysregulation of cellular mRNA stability in an HCV infection to another non-arthropod-borne member of the Flaviviridae, we assessed the abundance and stability of select cellular mRNAs in BVDV infected MDBK cells. As shown in Fig. 7A, the abundance of both FOS and JUN mRNAs was significantly upregulated (three to six fold) in a BVDV infection. Furthermore, the proteins encoded by these mRNAs were also upregulated in a BVDV infection (Fig. 7B) (and a similar increase in protein expression was seen in HCV infection (S4 Fig). As seen in Fig. 7C, the increase in abundance of the FOS and JUN mRNAs can largely be accounted for by the approximately five and three-fold significant increase in mRNA half-life in a BVDV infection, respectively. Importantly, XRN1 levels are not significantly changed during BVDV infection (S1B Fig). Thus, we conclude that dysregulation of cellular mRNA stability is a common occurrence in hepacivirus and pestivirus infections and may play a heretofore unforeseen role in the success of viral replication as well as viral-induced pathogenesis.

Bottom Line: We demonstrate that both Hepatitis C virus (HCV) and Bovine Viral Diarrhea virus (BVDV) contain regions in their 5' UTRs that stall and repress the enzymatic activity of the cellular 5'-3' exoribonuclease XRN1, resulting in dramatic changes in the stability of cellular mRNAs.In the context of HCV infection, we observed globally increased stability of mRNAs resulting in significant increases in abundance of normally short-lived mRNAs encoding a variety of relevant oncogenes and angiogenesis factors.These findings suggest that non-coding regions from multiple genera of the Flaviviridae interfere with XRN1 and impact post-transcriptional processes, causing global dysregulation of cellular gene expression which may promote cell growth and pathogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado, United States of America.

ABSTRACT
We demonstrate that both Hepatitis C virus (HCV) and Bovine Viral Diarrhea virus (BVDV) contain regions in their 5' UTRs that stall and repress the enzymatic activity of the cellular 5'-3' exoribonuclease XRN1, resulting in dramatic changes in the stability of cellular mRNAs. We used biochemical assays, virus infections, and transfection of the HCV and BVDV 5' untranslated regions in the absence of other viral gene products to directly demonstrate the existence and mechanism of this novel host-virus interaction. In the context of HCV infection, we observed globally increased stability of mRNAs resulting in significant increases in abundance of normally short-lived mRNAs encoding a variety of relevant oncogenes and angiogenesis factors. These findings suggest that non-coding regions from multiple genera of the Flaviviridae interfere with XRN1 and impact post-transcriptional processes, causing global dysregulation of cellular gene expression which may promote cell growth and pathogenesis.

Show MeSH
Related in: MedlinePlus