Protein–RNA tethering: The role of poly(C) binding protein 2 in poliovirus RNA replication

Spear, Allyn; Sharma, Nidhi; Flanegan, James B.

doi:10.1016/j.virol.2007.12.039

Cited by 28 publications

(39 citation statements)

References 46 publications

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“…It stabilizes the ␣-globin and collagen 1␣ (22) mRNAs in vivo through its interaction with three pyrimidine-rich patches within their 3= UTRs (68)(69)(70). PCBP2 is also implicated in the stabilization of viral RNAs and in the regulation of translation and/or replication and gene expression of several viruses, including poliovirus (30,(71)(72)(73), hepatitis A virus (HAV) (74), human papillomavirus (HPV) (75), vesicular stomatitis virus (VSV) (76), and HCV (27).…”

Section: Discussionmentioning

confidence: 99%

Norovirus Genome Circularization and Efficient Replication Are Facilitated by Binding of PCBP2 and hnRNP A1

López-Manríquez

Vashist

Ureña

et al. 2013

J Virol

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c Sequences and structures within the terminal genomic regions of plus-strand RNA viruses are targets for the binding of host proteins that modulate functions such as translation, RNA replication, and encapsidation. Using murine norovirus 1 (MNV-1), we describe the presence of long-range RNA-RNA interactions that were stabilized by cellular proteins. The proteins potentially responsible for the stabilization were selected based on their ability to bind the MNV-1 genome and/or having been reported to be involved in the stabilization of RNA-RNA interactions. Cell extracts were preincubated with antibodies against the selected proteins and used for coprecipitation reactions. Extracts treated with antibodies to poly(C) binding protein 2 (PCBP2) and heterogeneous nuclear ribonucleoprotein (hnRNP) A1 significantly reduced the 5=-3= interaction. Both PCBP2 and hnRNP A1 recombinant proteins stabilized the 5=-3= interactions and formed ribonucleoprotein complexes with the 5= and 3= ends of the MNV-1 genomic RNA. Mutations within the 3= complementary sequences (CS) that disrupt the 5=-3=-end interactions resulted in a significant reduction of the viral titer, suggesting that the integrity of the 3=-end sequence and/or the lack of complementarity with the 5= end is important for efficient virus replication. Small interfering RNA-mediated knockdown of PCBP2 or hnRNP A1 resulted in a reduction in virus yield, confirming a role for the observed interactions in efficient viral replication. PCBP2 and hnRNP A1 induced the circularization of MNV-1 RNA, as revealed by electron microscopy. This study provides evidence that PCBP2 and hnRNP A1 bind to the 5= and 3= ends of the MNV-1 viral RNA and contribute to RNA circularization, playing a role in the virus life cycle.

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Section: Discussionmentioning

confidence: 99%

Norovirus Genome Circularization and Efficient Replication Are Facilitated by Binding of PCBP2 and hnRNP A1

López-Manríquez

Vashist

Ureña

et al. 2013

J Virol

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“…Interactions between viral RNA and RNA-binding proteins play crucial roles in the positive-strand RNA virus life cycle, including regulating RNA synthesis, metabolism and stability (Spear et al, 2008). Y-box-binding protein 1 (YB-1), PTB, PABP, heterogeneous nuclear ribonucleoprotein Q, A1, and A2/B proteins have all been reported to bind specifically to the DENV 39 UTR (Paranjape & Harris, 2007;Yocupicio-Monroy et al, 2003).…”

Section: Discussionmentioning

confidence: 99%

Functional interaction between cellular p100 and the dengue virus 3' UTR

Lei

Huang

Hai

et al. 2010

Journal of General Virology

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Host factors interacting with the dengue virus (DENV) 39 UTR are involved in virus replication, but their roles remain poorly understood. We used RNA affinity capture and mass spectrometry to identify p100 as a host cellular protein associated with the DENV 39 UTR. By using RNA immunoprecipitation and confocal immunofluorescence analysis we demonstrated an interaction between p100 and the 39 UTR in DENV-infected cells. We identified the A4 region (the extensive stem-loop structure at the 39 end) as the binding site of p100 by studying deletion mutants. p100 knockdown specifically reduced the levels of viral RNA and viral protein in DENV-infected cells. Furthermore, downregulation of p100 reduced the expression of a heterologously expressed luciferase-39 UTR(DENV) mRNA in an A4-dependent manner, confirming the binding data and the effects of p100 knockdown on viral replication. These results provide evidence that p100 interacts with the 39 UTR of DENV and is required for normal DENV replication.

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“…(ii) PV1 pRzP23 was generated using PV1 pP23 plasmid by inserting a hammerhead ribozyme (Rz) downstream of the T7 promoter. PV1 RzP23 RNA transcribed from the pRzP23 plasmid contains a 5′ hammerhead ribozyme (Rz) which upon cleavage generates an authentic 5′ terminus which supports both (−) and (+) strand synthesis (Morasco et al, 2003;Sharma et al, 2005;Spear et al, 2008). …”

Section: Methodsmentioning

confidence: 99%

“…The 5′CL is a multifunctional element that is required for translation, (−) and (+) strand synthesis and VPg uridylylation (Barton et al, 2001;Gamarnik and Andino, 1998;Gamarnik and Andino, 2000;Murray et al, 2001;Ogram et al, 2010;Sharma et al, 2005;Spear et al, 2008;Sharma et al, 2009;Teterina et al, 2001;Vogt and Andino, 2010). The conserved cre hairpin structure in the 2C coding region of the RNA genome serves as the primary template for VPg uridylylation by 3D pol to form VPgpUpU (McKnight and Lemon, 1998;Goodfellow et al, 2000;Paul et al, 2000;Gerber et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

Amiloride inhibits the initiation of Coxsackievirus and poliovirus RNA replication by inhibiting VPg uridylylation

et al. 2014

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The mechanism of amiloride inhibition of Coxsackievirus B3 (CVB3) and poliovirus type 1 (PV1) RNA replication was investigated using membrane-associated RNA replication complexes. Amiloride was shown to inhibit viral RNA replication and VPgpUpU synthesis. However, the drug had no effect on polymerase elongation activity during either (−) strand or (+) strand synthesis. These findings indicated that amiloride inhibited the initiation of RNA synthesis by inhibiting VPg uridylylation. In addition, in silico binding studies showed that amiloride docks in the VPg binding site on the back of the viral RNA polymerase, 3Dpol. Since VPg binding at this site on PV1 3Dpol was previously shown to be required for VPg uridylylation, our results suggest that amiloride inhibits VPg binding to 3Dpol. In summary, our findings are consistent with a model in which amiloride inhibits VPgpUpU synthesis and viral RNA replication by competing with VPg for binding to 3Dpol.

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Protein–RNA tethering: The role of poly(C) binding protein 2 in poliovirus RNA replication

Cited by 28 publications

References 46 publications

Norovirus Genome Circularization and Efficient Replication Are Facilitated by Binding of PCBP2 and hnRNP A1

Norovirus Genome Circularization and Efficient Replication Are Facilitated by Binding of PCBP2 and hnRNP A1

Functional interaction between cellular p100 and the dengue virus 3' UTR

Amiloride inhibits the initiation of Coxsackievirus and poliovirus RNA replication by inhibiting VPg uridylylation

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