Viral RNA polymerase scanning and the gymnastics of Sendai virus RNA synthesis

Kolakofsky, Daniel; Mercier, Philippe Le; Iseni, Frédéric; Garcin, Dominique

doi:10.1016/j.virol.2003.10.031

Cited by 102 publications

(95 citation statements)

References 81 publications

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“…On the other hand, SeV (83) and Henipavirus would provide examples of the second scenario, with K D values in the M range. These findings would support a cartwheeling mechanism for the polymerase complex of Henipavirus, as already proposed for other Paramyxoviridae members (132,133). Whatever the mechanism by which the polymerase moves along the template, the present studies, by revealing that P is recruited onto the nucleocapsid template via the N TAIL -P XD interaction, designate this latter as a promising target for antiviral therapies.…”

Section: Structural Models Of Henipavirus N Tail -P Xd Complexes and supporting

confidence: 86%

Characterization of the Interactions between the Nucleoprotein and the Phosphoprotein of Henipavirus

Habchi

Blangy

Mamelli

et al. 2011

Journal of Biological Chemistry

165

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The Henipavirus genome is encapsidated by the nucleoprotein (N) within a helical nucleocapsid that recruits the polymerase complex via the phosphoprotein (P). In a previous study, we reported that in henipaviruses, the N-terminal domain of the phosphoprotein and the C-terminal domain of the nucleoprotein (N TAIL ) are both intrinsically disordered. Here we show that Henipavirus N TAIL domains are also disordered in the context of full-length nucleoproteins. We also report the cloning, purification, and characterization of the C-terminal X domains (P XD ) of Henipavirus phosphoproteins. Using isothermal titration calorimetry, we show that N TAIL and P XD form a 1:1 stoichiometric complex that is stable under NaCl concentrations as high as 1 M and has a K D in the M range. Using far-UV circular dichroism and nuclear magnetic resonance, we show that P XD triggers an increase in the ␣-helical content of N TAIL . Using fluorescence spectroscopy, we show that P XD has no impact on the chemical environment of a Trp residue introduced at position 527 of the Henipavirus N TAIL domain, thus arguing for the lack of stable contacts between the C termini of N TAIL and P XD . Finally, we present a tentative structural model of the N TAIL -P XD interaction in which a short, order-prone region of N TAIL (␣-MoRE; amino acids 473-493) adopts an ␣-helical conformation and is embedded between helices ␣2 and ␣3 of P XD , leading to a relatively small interface dominated by hydrophobic contacts. The present results provide the first detailed experimental characterization of the N-P interaction in henipaviruses and designate the N TAIL -P XD interaction as a valuable target for rational antiviral approaches.

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Section: Structural Models Of Henipavirus N Tail -P Xd Complexes and supporting

confidence: 86%

Characterization of the Interactions between the Nucleoprotein and the Phosphoprotein of Henipavirus

Habchi

Blangy

Mamelli

et al. 2011

Journal of Biological Chemistry

165

View full text Add to dashboard Cite

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“…SeV genomes contain six major proteins: NP, P, M, F, HN, and L expressed in a gradient with NP having the highest level of expression (40). Cells were infected with equal MOIs of SeV-52 or SeV-C for 3, 6, or 24 h. Quantitative analysis of mRNA extracted from these cells revealed that, at all time points, the cells infected with SeV-52 produced more viral NP mRNA than cells infected with SeV-C (Fig.…”

Section: Sev-c Produces More Activating Stimulus Than Sev-52mentioning

confidence: 99%

A Novel Role for Viral-Defective Interfering Particles in Enhancing Dendritic Cell Maturation

Yount

Kraus

Horvath

et al. 2006

The Journal of Immunology

109

152

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Dendritic cell (DC) maturation is a crucial event in the development of adaptive immune responses that confer long-lasting protection against reinfection with the same virus. Sendai virus strain Cantell has a particularly strong ability to mature DCs independently of type I IFNs and TLR signaling, currently the best-described pathways for the induction of DC maturation. In this study, we demonstrate that defective-interfering (DI) particles present in Sendai virus-Cantell stocks are required for its robust DC maturation ability. DI particles contain incomplete genomes that are unable to replicate unless the viral polymerase is supplied by coinfection with complete virus. Accordingly, the improvement in the virus-induced maturation of DCs provided by DI particles requires standard virus coinfection and likely results from increased production of dsRNA replication intermediaries. This unique ability of DI particles to stimulate DC maturation cannot be mimicked by simply increasing the dose of standard virus. Furthermore, viruses with weak DC maturation abilities can be converted into potent DC stimulators with the addition of DI particles, supporting a potential application for DI particles as a novel natural adjuvant for viral immunizations.

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“…This general model of polymerase organization has resulted from numerous studies on measles and closely related viruses (reviewed in Refs. [1][2][3][4][5][6]. The purpose of this note is to address a specific disagreement in the literature concerning the regions of measles N involved in binding the feet of the polymerase.…”

Section: Introductionmentioning

confidence: 99%

The feet of the measles virus polymerase bind the viral nucleocapsid protein at a single site

Yegambaram

Kingston

2010

Protein Science

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Measles virus has a single-stranded RNA genome that is organized into a helical complex by the viral N protein. The resulting structure is termed the nucleocapsid and is traversed by the viral polymerase during RNA synthesis. The P protein, the noncatalytic subunit of the polymerase, provides the ''legs and feet'' that allow the polymerase to walk along its protein-RNA template. The polymerase feet are very simple three-helix bundles, only 50 amino acids in size. Previously, we have shown that these feet grasp the viral N protein during movement by attaching to a short sequence (amino acids 487-503) within the disordered and surface-exposed tail of N, causing it to fold into a helix. The result is a weak-affinity complex with a short lifetime, which would allow the polymerase to take rapid steps forward. The structure of the complex was determined using X-ray crystallography. This simple model of binding was challenged by a paper in this journal, claiming that a downstream sequence in the tail of N (amino acids 517-525) was also critical for the association. Its presence was reported to enhance the overall affinity of the polymerase feet for N by three orders of magnitude. We have, therefore, examined binding of the polymerase foot domain to amino acids 477-525 of N using quantitative biophysical techniques, and compared the results to our previous binding studies, performed using amino acids 477-505 of N. We find no evidence that the sequence downstream of amino acid 505 influences binding, validating the original single-site binding model.

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Viral RNA polymerase scanning and the gymnastics of Sendai virus RNA synthesis

Cited by 102 publications

References 81 publications

Characterization of the Interactions between the Nucleoprotein and the Phosphoprotein of Henipavirus

Characterization of the Interactions between the Nucleoprotein and the Phosphoprotein of Henipavirus

A Novel Role for Viral-Defective Interfering Particles in Enhancing Dendritic Cell Maturation

The feet of the measles virus polymerase bind the viral nucleocapsid protein at a single site

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