The non-structural protein Nsp10 of mouse hepatitis virus binds zinc ions and nucleic acids

Nele Matthes; Jeroen R. Mesters; Bruno Coutard; Bruno Canard; Eric J. Snijder; Ralf Moll; Rolf Hilgenfeld

doi:10.1016/j.febslet.2006.06.061

The non-structural protein Nsp10 of mouse hepatitis virus binds zinc ions and nucleic acids

Nele Matthes, Jeroen R. Mesters, Bruno Coutard, Bruno Canard, Eric J. Snijder, Ralf Moll, Rolf Hilgenfeld^*

^*Korrespondierende/r Autor/-in für diese Arbeit

Institut für Biochemie

35 Zitate (Scopus)

Abstract

The non-structural protein Nsp10 of coronaviruses is a small cleavage product of the viral replicase polyprotein that has been implicated in RNA synthesis. Nsp10 of mouse hepatitis virus (MHV) displays an apparent molecular mass of 13-16 kDa in reducing SDS-PAGE and analytical gel filtration, while dynamic light scattering suggests the existence of oligomeric forms. Atomic absorption spectroscopy reveals two metal ions per Nsp10 monomer, with a preference for Zn²⁺ over Fe^2+/3+ and Co²⁺. These are probably bound by two Zn-finger-like motifs. Moreover, MHV Nsp10 interacts with tRNA, single-stranded RNA, double-stranded DNA and, to a lesser extent, single-stranded DNA as shown by gel-shift experiments. The K_d for tRNA is 2.1 ± 0.2 μM.

Originalsprache	Englisch
Zeitschrift	FEBS Letters
Jahrgang	580
Ausgabenummer	17
Seiten (von - bis)	4143-4149
Seitenumfang	7
ISSN	0014-5793
DOIs	https://doi.org/10.1016/j.febslet.2006.06.061
Publikationsstatus	Veröffentlicht - 24.07.2006

Fördermittel

This work was funded by the VIZIER project of the European Commission (contract No. LSHG-CT-2004-511960; www.vizier-europe.org ) and, in part, by the Sino-European Project on SARS Diagnostics and Antivirals (SEPSDA, SP22-CT-2004-003831; www.sepsda.info ). R.H. thanks the Fonds der Chemischen Industrie for support. E.S. is grateful to Volker Thiel and Stuart Siddell for providing recombinant vaccinia virus vMHV-inf-1, and acknowledges the general and/or technical support of Clara Posthuma, Linda van der Zanden and Alexander Gorbalenya.

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Forschungsschwerpunkt: Infektion und Entzündung - Zentrum für Infektions- und Entzündungsforschung Lübeck (ZIEL)

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10.1016/j.febslet.2006.06.061

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title = "The non-structural protein Nsp10 of mouse hepatitis virus binds zinc ions and nucleic acids",

abstract = "The non-structural protein Nsp10 of coronaviruses is a small cleavage product of the viral replicase polyprotein that has been implicated in RNA synthesis. Nsp10 of mouse hepatitis virus (MHV) displays an apparent molecular mass of 13-16 kDa in reducing SDS-PAGE and analytical gel filtration, while dynamic light scattering suggests the existence of oligomeric forms. Atomic absorption spectroscopy reveals two metal ions per Nsp10 monomer, with a preference for Zn2+ over Fe2+/3+ and Co2+. These are probably bound by two Zn-finger-like motifs. Moreover, MHV Nsp10 interacts with tRNA, single-stranded RNA, double-stranded DNA and, to a lesser extent, single-stranded DNA as shown by gel-shift experiments. The Kd for tRNA is 2.1 ± 0.2 μM.",

author = "Nele Matthes and Mesters, \{Jeroen R.\} and Bruno Coutard and Bruno Canard and Snijder, \{Eric J.\} and Ralf Moll and Rolf Hilgenfeld",

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T1 - The non-structural protein Nsp10 of mouse hepatitis virus binds zinc ions and nucleic acids

AU - Matthes, Nele

AU - Mesters, Jeroen R.

AU - Coutard, Bruno

AU - Canard, Bruno

AU - Snijder, Eric J.

AU - Moll, Ralf

AU - Hilgenfeld, Rolf

PY - 2006/7/24

Y1 - 2006/7/24

N2 - The non-structural protein Nsp10 of coronaviruses is a small cleavage product of the viral replicase polyprotein that has been implicated in RNA synthesis. Nsp10 of mouse hepatitis virus (MHV) displays an apparent molecular mass of 13-16 kDa in reducing SDS-PAGE and analytical gel filtration, while dynamic light scattering suggests the existence of oligomeric forms. Atomic absorption spectroscopy reveals two metal ions per Nsp10 monomer, with a preference for Zn2+ over Fe2+/3+ and Co2+. These are probably bound by two Zn-finger-like motifs. Moreover, MHV Nsp10 interacts with tRNA, single-stranded RNA, double-stranded DNA and, to a lesser extent, single-stranded DNA as shown by gel-shift experiments. The Kd for tRNA is 2.1 ± 0.2 μM.

AB - The non-structural protein Nsp10 of coronaviruses is a small cleavage product of the viral replicase polyprotein that has been implicated in RNA synthesis. Nsp10 of mouse hepatitis virus (MHV) displays an apparent molecular mass of 13-16 kDa in reducing SDS-PAGE and analytical gel filtration, while dynamic light scattering suggests the existence of oligomeric forms. Atomic absorption spectroscopy reveals two metal ions per Nsp10 monomer, with a preference for Zn2+ over Fe2+/3+ and Co2+. These are probably bound by two Zn-finger-like motifs. Moreover, MHV Nsp10 interacts with tRNA, single-stranded RNA, double-stranded DNA and, to a lesser extent, single-stranded DNA as shown by gel-shift experiments. The Kd for tRNA is 2.1 ± 0.2 μM.

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The non-structural protein Nsp10 of mouse hepatitis virus binds zinc ions and nucleic acids

Abstract

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