Structural disorder within Henipavirus nucleoprotein and phosphoprotein: from predictions to experimental assessment.

PloS One
Johnny HabchiSonia Longhi

Abstract

Henipaviruses are newly emerged viruses within the Paramyxoviridae family. Their negative-strand RNA genome is packaged by the nucleoprotein (N) within alpha-helical nucleocapsid that recruits the polymerase complex made of the L protein and the phosphoprotein (P). To date structural data on Henipaviruses are scarce, and their N and P proteins have never been characterized so far. Using both computational and experimental approaches we herein show that Henipaviruses N and P proteins possess large intrinsically disordered regions. By combining several disorder prediction methods, we show that the N-terminal domain of P (PNT) and the C-terminal domain of N (NTAIL) are both mostly disordered, although they contain short order-prone segments. We then report the cloning, the bacterial expression, purification and characterization of Henipavirus PNT and NTAIL domains. By combining gel filtration, dynamic light scattering, circular dichroism and nuclear magnetic resonance, we show that both NTAIL and PNT belong to the premolten globule sub-family within the class of intrinsically disordered proteins. This study is the first reported experimental characterization of Henipavirus P and N proteins. The evidence that their respective N-ter...Continue Reading

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Citations

Jun 11, 2014·Acta Crystallographica. Section D, Biological Crystallography·David BlocquelSonia Longhi
May 16, 2014·Chemical Reviews·Bin XueSonia Longhi
Aug 12, 2014·Nature Structural & Molecular Biology·Filip YabukarskiMarc Jamin
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Jun 14, 2015·FEBS Letters·Sonia Longhi
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Datasets Mentioned

BETA
ABM64790.1
ABY61984.1
BAI60055.1
AAB06278.1

Methods Mentioned

BETA
gel filtration
Affinity
gel
size-exclusion chromatography
circular dichroism
dynamic light scattering
NMR
PCR
gel filtrations
protein assay

Software Mentioned

MoRES
IDP
ClustalW
RONN
CDSSTR
Bruker Topspin
Emboss
MeDor metaserver
EXPASY
Protscale

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