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Characterization of the interactions between the nucleoprotein and the phosphoprotein of Henipavirus.
- Source :
-
The Journal of biological chemistry [J Biol Chem] 2011 Apr 15; Vol. 286 (15), pp. 13583-602. Date of Electronic Publication: 2011 Feb 11. - Publication Year :
- 2011
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Abstract
- 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 μM range. Using far-UV circular dichroism and nuclear magnetic resonance, we show that P(XD) triggers an increase in the α-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) (α-MoRE; amino acids 473-493) adopts an α-helical conformation and is embedded between helices α2 and α3 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.
- Subjects :
- Henipavirus genetics
Nucleoproteins genetics
Phosphoproteins genetics
Protein Structure, Quaternary
Protein Structure, Secondary
Protein Structure, Tertiary
Recombinant Proteins chemistry
Recombinant Proteins genetics
Viral Proteins genetics
Henipavirus chemistry
Models, Molecular
Nucleoproteins chemistry
Phosphoproteins chemistry
Viral Proteins chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1083-351X
- Volume :
- 286
- Issue :
- 15
- Database :
- MEDLINE
- Journal :
- The Journal of biological chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 21317293
- Full Text :
- https://doi.org/10.1074/jbc.M111.219857