Interaction of poly(rC)-binding protein 2 domains KH1 and KH3 with coxsackievirus RNA

Zell, Roland; Ihle, Yvonne; Effenberger, Madlen; Seitz, Simone; Wutzler, Peter; Görlach, Matthias

doi:10.1016/j.bbrc.2008.09.156

Cited by 13 publications

(16 citation statements)

References 21 publications

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“…The predominantly nuclear PCBP2 binds to both stem-loop IV (S-L IV) and stem-loop I (S-L I) of the 5′-NCR within the poliovirus and CVB3 genomic RNA (Blyn et al, 1995, 1996; Leffers et al, 1995; Gamarnik and Andino, 1997; Parsley et al, 1997; Zell et al, 2008a,b; Sean et al, 2009). S-L IV is located in the central portion of Type I IRES elements and alterations to the nucleic acid sequence identified as important for PCBP2 association decrease poliovirus translation in vitro (Blyn et al, 1995).…”

Section: Nuclear-resident Proteins Are Hijacked For Picornavirus Tranmentioning

confidence: 99%

Picornaviruses and nuclear functions: targeting a cellular compartment distinct from the replication site of a positive-strand RNA virus

Flather

Semler

2015

Front. Microbiol.

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The compartmentalization of DNA replication and gene transcription in the nucleus and protein production in the cytoplasm is a defining feature of eukaryotic cells. The nucleus functions to maintain the integrity of the nuclear genome of the cell and to control gene expression based on intracellular and environmental signals received through the cytoplasm. The spatial separation of the major processes that lead to the expression of protein-coding genes establishes the necessity of a transport network to allow biomolecules to translocate between these two regions of the cell. The nucleocytoplasmic transport network is therefore essential for regulating normal cellular functioning. The Picornaviridae virus family is one of many viral families that disrupt the nucleocytoplasmic trafficking of cells to promote viral replication. Picornaviruses contain positive-sense, single-stranded RNA genomes and replicate in the cytoplasm of infected cells. As a result of the limited coding capacity of these viruses, cellular proteins are required by these intracellular parasites for both translation and genomic RNA replication. Being of messenger RNA polarity, a picornavirus genome can immediately be translated upon entering the cell cytoplasm. However, the replication of viral RNA requires the activity of RNA-binding proteins, many of which function in host gene expression, and are consequently localized to the nucleus. As a result, picornaviruses disrupt nucleocytoplasmic trafficking to exploit protein functions normally localized to a different cellular compartment from which they translate their genome to facilitate efficient replication. Furthermore, picornavirus proteins are also known to enter the nucleus of infected cells to limit host-cell transcription and down-regulate innate antiviral responses. The interactions of picornavirus proteins and host-cell nuclei are extensive, required for a productive infection, and are the focus of this review.

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Section: Nuclear-resident Proteins Are Hijacked For Picornavirus Tranmentioning

confidence: 99%

Picornaviruses and nuclear functions: targeting a cellular compartment distinct from the replication site of a positive-strand RNA virus

Flather

Semler

2015

Front. Microbiol.

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“…PCBP2's three KH domains all bind to the apical region of dIV in Type 1 IRESs, with the strongest interactions involving dIVb and dIVc (3,4,13,24). With the exception of d10c, the structure of the element of the CDV IRES to which PCBP2 binds differs significantly from dIV.…”

Section: Resultsmentioning

confidence: 99%

“…27–34,53). PCBP2 binds specifically via its KH1 domain to an exposed internal C-rich loop in the apical subdomain IVc of Type 1 IRESs (3,4,13,24) and to the CDV IRES by interaction with the closely related d10c, protecting an analogous pyrimidine-rich loop from NMIA modification (Figure 1C). This site of interaction is critical for the function of the CDV IRES (this report) and for canonical Type 1 IRESs (3,4).…”

Section: Discussionmentioning

confidence: 99%

“…Although KH3 has a high degree of mobility in solution (21), it is in close proximity to KH2 in CDV IRES-bound PCBP2, as also noted for PCBP2 bound to PV and EV71 IRESs (13). However, whereas the KH3 binding site on these and other canonical Type 1 IRESs is centered on the pyrimidine-rich loop motif at the apex of domain IVb (3,4,13,24), the KH3 binding site on the CDV IRES is located centrally in d10b. Moreover, the orientation of KH3 relative to KH2 is flipped, so that when PCBP2 is bound to e.g.…”

Section: Discussionmentioning

confidence: 99%

“…Binding of RNA involves a hydrophobic cleft formed on one side by α 1 , α 2 and the intervening conserved GXXG loop, which interacts with the RNA backbone and orients four bases to the other side, which is formed by the β-sheet and a variable loop (22,23). PCBP2 engages in intricate interactions with the apical region of dIV of Type 1 IRESs, including a major interaction of KH1 with the C-rich loop in dIVc (3,4,13,24). Various functions have been proposed for PCBP2 and other ITAFs in promoting IRES-mediated initiation, including interaction with and recruitment of initiation factors, other ITAFs or the 40S ribosomal subunit to the IRES, and stabilization of IRES structure in an active conformation (25).…”

Section: Introductionmentioning

confidence: 99%

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PCBP2 enables the cadicivirus IRES to exploit the function of a conserved GRNA tetraloop to enhance ribosomal initiation complex formation

2016

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The cadicivirus IRES diverges structurally from canonical Type 1 IRESs (e.g. poliovirus) but nevertheless also contains an essential GNRA tetraloop in a subdomain (d10c) that is homologous to poliovirus dIVc. In addition to canonical initiation factors, the canonical Type 1 and divergent cadicivirus IRESs require the same IRES trans-acting factor, poly(C)-binding protein 2 (PCBP2). PCBP2 has three KH domains and binds poliovirus IRES domain dIV in the vicinity of the tetraloop. How PCBP2 binds the cadicivirus IRES, and the roles of PCBP2 and the tetraloop in Type 1 IRES function are unknown. Here, directed hydroxyl radical probing showed that KH1 also binds near the cadicivirus tetraloop. KH2 and KH3 bind adjacently to an IRES subdomain (d10b) that is unrelated to dIV, with KH3 in an inverted orientation. KH3 is critical for PCBP2's binding to this IRES whereas KH1 is essential for PCBP2's function in promoting initiation. PCBP2 enforced the wild-type structure of d10c when it contained minor destabilizing substitutions, exposing the tetraloop. Strikingly, PCBP2 enhanced initiation on mutant IRESs that retained consensus GNRA tetraloops, whereas mutants with divergent sequences did not respond to PCBP2. These studies show that PCBP2 enables the IRES to exploit the GNRA tetraloop to enhance initiation.

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The mechanism of translation initiation on Type 1 picornavirus IRESs

Sweeney

Abaeva

Pestova

et al. 2013

EMBO J

141

206

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Picornavirus Type 1 IRESs comprise five principal domains (dII-dVI). Whereas dV binds eIF4G, a conserved AUG in dVI was suggested to stimulate attachment of 43S ribosomal preinitiation complexes, which then scan to the initiation codon. Initiation on Type 1 IRESs also requires IRES trans-acting factors (ITAFs), and several candidates have been proposed. Here, we report the in vitro reconstitution of initiation on three Type 1 IRESs: poliovirus (PV), enterovirus 71 (EV71), and bovine enterovirus (BEV). All of them require eIF2, eIF3, eIF4A, eIF4G, eIF4B, eIF1A, and a single ITAF, poly(C) binding protein 2 (PCBP2). In each instance, initiation starts with binding of eIF4G/eIF4A. Subsequent recruitment of 43S complexes strictly requires direct interaction of their eIF3 constituent with eIF4G. The following events can differ between IRESs, depending on the stability of dVI. If it is unstructured (BEV), all ribosomes scan through dVI to the initiation codon, requiring eIF1 to bypass its AUG. If it is structured (PV, EV71), most initiation events occur without inspection of dVI, implying that its AUG does not determine ribosomal attachment.

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Interaction of poly(rC)-binding protein 2 domains KH1 and KH3 with coxsackievirus RNA

Cited by 13 publications

References 21 publications

Picornaviruses and nuclear functions: targeting a cellular compartment distinct from the replication site of a positive-strand RNA virus

Picornaviruses and nuclear functions: targeting a cellular compartment distinct from the replication site of a positive-strand RNA virus

PCBP2 enables the cadicivirus IRES to exploit the function of a conserved GRNA tetraloop to enhance ribosomal initiation complex formation

The mechanism of translation initiation on Type 1 picornavirus IRESs

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