A Conserved Basic Patch and Central Kink in the Nipah Virus Phosphoprotein Multimerization Domain Are Essential for Polymerase Function

Jessica F Bruhn; Anne L Hotard; Christina F Spiropoulou; Michael K Lo; Erica Ollmann Saphire

doi:10.1016/j.str.2019.01.012

A Conserved Basic Patch and Central Kink in the Nipah Virus Phosphoprotein Multimerization Domain Are Essential for Polymerase Function

Structure. 2019 Apr 2;27(4):660-668.e4. doi: 10.1016/j.str.2019.01.012. Epub 2019 Feb 21.

Authors

Jessica F Bruhn¹, Anne L Hotard², Christina F Spiropoulou², Michael K Lo³, Erica Ollmann Saphire⁴

Affiliations

¹ Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA 92037, USA.
² Viral Special Pathogens Branch, Centers for Disease Control and Prevention, Atlanta, GA 30329, USA.
³ Viral Special Pathogens Branch, Centers for Disease Control and Prevention, Atlanta, GA 30329, USA. Electronic address: mko2@cdc.gov.
⁴ Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA 92037, USA; The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037, USA. Electronic address: erica@scripps.edu.

Abstract

Nipah virus is a highly lethal zoonotic pathogen found in Southeast Asia that has caused human encephalitis outbreaks with 40%-70% mortality. NiV encodes its own RNA-dependent RNA polymerase within the large protein, L. Efficient polymerase activity requires the phosphoprotein, P, which tethers L to its template, the viral nucleocapsid. P is a multifunctional protein with modular domains. The central P multimerization domain is composed of a long, tetrameric coiled coil. We investigated the importance of structural features found in this domain for polymerase function using a newly constructed NiV bicistronic minigenome assay. We identified a conserved basic patch and central kink in the coiled coil that are important for polymerase function, with R555 being absolutely essential. This basic patch and central kink are conserved in the related human pathogens measles and mumps viruses, suggesting that this mechanism may be conserved.

Keywords: Nipah virus; RNA-dependent RNA polymerase; X-ray crystallography; coiled coil; differential scanning calorimetry; henipavirus; minigenome; oligomerization; phosphoprotein.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Amino Acid Sequence
Binding Sites
Cloning, Molecular
Conserved Sequence
Crystallography, X-Ray
DNA-Directed RNA Polymerases / chemistry*
DNA-Directed RNA Polymerases / genetics
DNA-Directed RNA Polymerases / metabolism
Escherichia coli / genetics
Escherichia coli / metabolism
Gene Expression
Genetic Vectors / chemistry
Genetic Vectors / metabolism
Genome, Viral*
Humans
Measles virus / chemistry
Measles virus / enzymology
Measles virus / genetics
Models, Molecular
Mumps virus / chemistry
Mumps virus / enzymology
Mumps virus / genetics
Nipah Virus / chemistry*
Nipah Virus / enzymology
Nipah Virus / genetics
Phosphoproteins / chemistry*
Phosphoproteins / genetics
Phosphoproteins / metabolism
Protein Binding
Protein Conformation, alpha-Helical
Protein Interaction Domains and Motifs
Protein Multimerization
RNA-Dependent RNA Polymerase / chemistry*
RNA-Dependent RNA Polymerase / genetics
RNA-Dependent RNA Polymerase / metabolism
Recombinant Proteins / chemistry
Recombinant Proteins / genetics
Recombinant Proteins / metabolism
Sequence Alignment
Sequence Homology, Amino Acid
Viral Proteins / chemistry*
Viral Proteins / genetics
Viral Proteins / metabolism

Substances

P protein, Nipah virus
Phosphoproteins
Recombinant Proteins
Viral Proteins
L protein, Nipah virus
RNA-Dependent RNA Polymerase
DNA-Directed RNA Polymerases

Abstract

Publication types

MeSH terms

Substances

Grants and funding