Structural insights into phosphoprotein chaperoning of nucleoprotein in measles virus

Sergey Guryanov, Lassi Juho Petteri Liljeroos, Prasad Kasaragod, Tommi Antero Kajander, Sarah Jane Butcher

Tutkimustuotos: KonferenssimateriaalitPosteri

Kuvaus

Measles virus is an important, highly contagious, human pathogen. The nucleoprotein N binds only to viral genomic RNA and forms the helical ribonucleocapsid that serves as a template for viral replication. We address how N is regulated by another protein, the phosphoprotein, P, to prevent newly synthesized N from binding to cellular RNA. Here, we pulled down an N01-408 fragment lacking most of its C-terminal tail domain by several affinity-tagged, N-terminal, P fragments to map the N0-binding region of P to the first 48 amino acids. We showed biochemically and using P mutants the importance of the hydrophobic interactions for the binding. We fused an N0 binding peptide, P1-48, to the C-terminus of an N021-408 fragment lacking both the N-terminal peptide and the C-terminal tail of N protein to reconstitute and crystallize the N0-P complex. We solved the X-ray structure of the resulting N0-P chimeric protein at 2.7 Å resolution. The structure reveals the molecular details of the conserved N0-P interface and explains how P chaperones N0 preventing both self-assembly of N0 and its binding to RNA. We compare the structure of an N0-P complex to atomic model of helical ribonucleocapsid. We thus propose a model how P may help to start viral RNA synthesis. Our results provide a new insight into mechanisms of paramyxovirus replication. New data on the mechanisms of phosphoprotein chaperone action allows better understanding of the virus genome replication and nucleocapsid assembly. We describe a conserved structural interface for the N-P interaction which could be a target for drug development not only to treat measles but also potentially other paramyxovirus diseases.
Alkuperäiskielienglanti
Sivut44
Sivumäärä1
TilaJulkaistu - 2017
Tapahtuma2nd Annual Users Meeting of iNEXT - International Best Western Hotel, Brno, Tšekki
Kesto: 22 toukokuuta 201724 toukokuuta 2017
http://inext.ceitec.eu/

Konferenssi

Konferenssi2nd Annual Users Meeting of iNEXT
MaaTšekki
KaupunkiBrno
Ajanjakso22/05/201724/05/2017
www-osoite

Tieteenalat

  • 1182 Biokemia, solu- ja molekyylibiologia

Lainaa tätä

Guryanov, S., Liljeroos, L. J. P., Kasaragod, P., Kajander, T. A., & Butcher, S. J. (2017). Structural insights into phosphoprotein chaperoning of nucleoprotein in measles virus. 44. Posterin esittämispaikka: 2nd Annual Users Meeting of iNEXT, Brno, Tšekki.
Guryanov, Sergey ; Liljeroos, Lassi Juho Petteri ; Kasaragod, Prasad ; Kajander, Tommi Antero ; Butcher, Sarah Jane. / Structural insights into phosphoprotein chaperoning of nucleoprotein in measles virus. Posterin esittämispaikka: 2nd Annual Users Meeting of iNEXT, Brno, Tšekki.1 Sivumäärä
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title = "Structural insights into phosphoprotein chaperoning of nucleoprotein in measles virus",
abstract = "Measles virus is an important, highly contagious, human pathogen. The nucleoprotein N binds only to viral genomic RNA and forms the helical ribonucleocapsid that serves as a template for viral replication. We address how N is regulated by another protein, the phosphoprotein, P, to prevent newly synthesized N from binding to cellular RNA. Here, we pulled down an N01-408 fragment lacking most of its C-terminal tail domain by several affinity-tagged, N-terminal, P fragments to map the N0-binding region of P to the first 48 amino acids. We showed biochemically and using P mutants the importance of the hydrophobic interactions for the binding. We fused an N0 binding peptide, P1-48, to the C-terminus of an N021-408 fragment lacking both the N-terminal peptide and the C-terminal tail of N protein to reconstitute and crystallize the N0-P complex. We solved the X-ray structure of the resulting N0-P chimeric protein at 2.7 {\AA} resolution. The structure reveals the molecular details of the conserved N0-P interface and explains how P chaperones N0 preventing both self-assembly of N0 and its binding to RNA. We compare the structure of an N0-P complex to atomic model of helical ribonucleocapsid. We thus propose a model how P may help to start viral RNA synthesis. Our results provide a new insight into mechanisms of paramyxovirus replication. New data on the mechanisms of phosphoprotein chaperone action allows better understanding of the virus genome replication and nucleocapsid assembly. We describe a conserved structural interface for the N-P interaction which could be a target for drug development not only to treat measles but also potentially other paramyxovirus diseases.",
keywords = "1182 Biochemistry, cell and molecular biology",
author = "Sergey Guryanov and Liljeroos, {Lassi Juho Petteri} and Prasad Kasaragod and Kajander, {Tommi Antero} and Butcher, {Sarah Jane}",
note = "Poster; 2nd Annual Users Meeting of iNEXT ; Conference date: 22-05-2017 Through 24-05-2017",
year = "2017",
language = "English",
pages = "44",
url = "http://inext.ceitec.eu/",

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Guryanov, S, Liljeroos, LJP, Kasaragod, P, Kajander, TA & Butcher, SJ 2017, 'Structural insights into phosphoprotein chaperoning of nucleoprotein in measles virus', 2nd Annual Users Meeting of iNEXT, Brno, Tšekki, 22/05/2017 - 24/05/2017 Sivut 44.

Structural insights into phosphoprotein chaperoning of nucleoprotein in measles virus. / Guryanov, Sergey; Liljeroos, Lassi Juho Petteri; Kasaragod, Prasad; Kajander, Tommi Antero; Butcher, Sarah Jane.

2017. 44 Posterin esittämispaikka: 2nd Annual Users Meeting of iNEXT, Brno, Tšekki.

Tutkimustuotos: KonferenssimateriaalitPosteri

TY - CONF

T1 - Structural insights into phosphoprotein chaperoning of nucleoprotein in measles virus

AU - Guryanov, Sergey

AU - Liljeroos, Lassi Juho Petteri

AU - Kasaragod, Prasad

AU - Kajander, Tommi Antero

AU - Butcher, Sarah Jane

N1 - Poster

PY - 2017

Y1 - 2017

N2 - Measles virus is an important, highly contagious, human pathogen. The nucleoprotein N binds only to viral genomic RNA and forms the helical ribonucleocapsid that serves as a template for viral replication. We address how N is regulated by another protein, the phosphoprotein, P, to prevent newly synthesized N from binding to cellular RNA. Here, we pulled down an N01-408 fragment lacking most of its C-terminal tail domain by several affinity-tagged, N-terminal, P fragments to map the N0-binding region of P to the first 48 amino acids. We showed biochemically and using P mutants the importance of the hydrophobic interactions for the binding. We fused an N0 binding peptide, P1-48, to the C-terminus of an N021-408 fragment lacking both the N-terminal peptide and the C-terminal tail of N protein to reconstitute and crystallize the N0-P complex. We solved the X-ray structure of the resulting N0-P chimeric protein at 2.7 Å resolution. The structure reveals the molecular details of the conserved N0-P interface and explains how P chaperones N0 preventing both self-assembly of N0 and its binding to RNA. We compare the structure of an N0-P complex to atomic model of helical ribonucleocapsid. We thus propose a model how P may help to start viral RNA synthesis. Our results provide a new insight into mechanisms of paramyxovirus replication. New data on the mechanisms of phosphoprotein chaperone action allows better understanding of the virus genome replication and nucleocapsid assembly. We describe a conserved structural interface for the N-P interaction which could be a target for drug development not only to treat measles but also potentially other paramyxovirus diseases.

AB - Measles virus is an important, highly contagious, human pathogen. The nucleoprotein N binds only to viral genomic RNA and forms the helical ribonucleocapsid that serves as a template for viral replication. We address how N is regulated by another protein, the phosphoprotein, P, to prevent newly synthesized N from binding to cellular RNA. Here, we pulled down an N01-408 fragment lacking most of its C-terminal tail domain by several affinity-tagged, N-terminal, P fragments to map the N0-binding region of P to the first 48 amino acids. We showed biochemically and using P mutants the importance of the hydrophobic interactions for the binding. We fused an N0 binding peptide, P1-48, to the C-terminus of an N021-408 fragment lacking both the N-terminal peptide and the C-terminal tail of N protein to reconstitute and crystallize the N0-P complex. We solved the X-ray structure of the resulting N0-P chimeric protein at 2.7 Å resolution. The structure reveals the molecular details of the conserved N0-P interface and explains how P chaperones N0 preventing both self-assembly of N0 and its binding to RNA. We compare the structure of an N0-P complex to atomic model of helical ribonucleocapsid. We thus propose a model how P may help to start viral RNA synthesis. Our results provide a new insight into mechanisms of paramyxovirus replication. New data on the mechanisms of phosphoprotein chaperone action allows better understanding of the virus genome replication and nucleocapsid assembly. We describe a conserved structural interface for the N-P interaction which could be a target for drug development not only to treat measles but also potentially other paramyxovirus diseases.

KW - 1182 Biochemistry, cell and molecular biology

M3 - Poster

SP - 44

ER -

Guryanov S, Liljeroos LJP, Kasaragod P, Kajander TA, Butcher SJ. Structural insights into phosphoprotein chaperoning of nucleoprotein in measles virus. 2017. Posterin esittämispaikka: 2nd Annual Users Meeting of iNEXT, Brno, Tšekki.