A Zinc Ion Controls Assembly and Stability of the Major Capsid Protein of Rotavirus

The recent determination of the crystal structure of VP6, the major capsid protein of rotavirus, revealed a trimer containing a central zinc ion coordinated by histidine 153 from each of the three subunits. The role of the zinc ion in the functions of VP6 was investigated by site-directed mutagenesi...

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Päätekijät: Erk, Inge, Huet, Jean-Claude, Duarte, Mariela, Duquerroy, Stéphane, Rey, Felix, Cohen, Jean, Lepault, Jean
Aineistotyyppi: Artikkeli
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Julkaistu: American Society for Microbiology 2003
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Structure and Assembly
Linkit:https://ncbi.nlm.nih.gov/pmc/articles/PMC149495/
https://ncbi.nlm.nih.gov/pubmed/12610135
https://ncbi.nlm.nih.govhttp://dx.doi.org/10.1128/JVI.77.6.3595-3601.2003
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spelling pubmed-1494952003-04-02 A Zinc Ion Controls Assembly and Stability of the Major Capsid Protein of Rotavirus Erk, Inge Huet, Jean-Claude Duarte, Mariela Duquerroy, Stéphane Rey, Felix Cohen, Jean Lepault, Jean J Virol Structure and Assembly The recent determination of the crystal structure of VP6, the major capsid protein of rotavirus, revealed a trimer containing a central zinc ion coordinated by histidine 153 from each of the three subunits. The role of the zinc ion in the functions of VP6 was investigated by site-directed mutagenesis. The mutation of histidine 153 into a serine (H153S and H153S/S339H) did not prevent the formation of VP6 trimers. At pH <7.0, about the pK of histidine, wild-type and mutated VP6 proteins display similar properties, giving rise to identical tubular and spherical assemblies. However, at pH >7.0, histidine 153 mutant proteins did not assemble into the characteristic 45-nm-diameter tubes, in contrast to wild-type VP6. These observations showed that under conditions in which histidine residues are not charged, the properties of VP6 depended on the presence of the centrally coordinated zinc atom in the trimer. Indeed, wild-type VP6 depleted of the zinc ion by a high concentration (100 mM) of a metal-chelating agent behaved like the H153 mutant proteins. The susceptibility of wild-type VP6 to proteases is greatly increased in the absence of zinc. NH(2)-terminal sequencing of the proteolytic fragments showed that they all contained the β-sheet-rich VP6 head domain, which appeared to be less sensitive to protease activity than the α-helical basal domain. Finally, the mutant proteins assembled well on cores, as demonstrated by both electron microscopy and rescue of transcriptase activity. Zinc is thus not necessary for the transcription activity. All of these observations suggest that, in solution, VP6 trimers present a structural flexibility that is controlled by the presence of a zinc ion. American Society for Microbiology 2003-03 /pmc/articles/PMC149495/ /pubmed/12610135 http://dx.doi.org/10.1128/JVI.77.6.3595-3601.2003 Text en Copyright © 2003, American Society for Microbiology
institution US National Library of Medicine
collection PubMed Central
language en
format Article
topic Structure and Assembly
spellingShingle Structure and Assembly
Erk, Inge
Huet, Jean-Claude
Duarte, Mariela
Duquerroy, Stéphane
Rey, Felix
Cohen, Jean
Lepault, Jean
A Zinc Ion Controls Assembly and Stability of the Major Capsid Protein of Rotavirus
description The recent determination of the crystal structure of VP6, the major capsid protein of rotavirus, revealed a trimer containing a central zinc ion coordinated by histidine 153 from each of the three subunits. The role of the zinc ion in the functions of VP6 was investigated by site-directed mutagenesis. The mutation of histidine 153 into a serine (H153S and H153S/S339H) did not prevent the formation of VP6 trimers. At pH <7.0, about the pK of histidine, wild-type and mutated VP6 proteins display similar properties, giving rise to identical tubular and spherical assemblies. However, at pH >7.0, histidine 153 mutant proteins did not assemble into the characteristic 45-nm-diameter tubes, in contrast to wild-type VP6. These observations showed that under conditions in which histidine residues are not charged, the properties of VP6 depended on the presence of the centrally coordinated zinc atom in the trimer. Indeed, wild-type VP6 depleted of the zinc ion by a high concentration (100 mM) of a metal-chelating agent behaved like the H153 mutant proteins. The susceptibility of wild-type VP6 to proteases is greatly increased in the absence of zinc. NH(2)-terminal sequencing of the proteolytic fragments showed that they all contained the β-sheet-rich VP6 head domain, which appeared to be less sensitive to protease activity than the α-helical basal domain. Finally, the mutant proteins assembled well on cores, as demonstrated by both electron microscopy and rescue of transcriptase activity. Zinc is thus not necessary for the transcription activity. All of these observations suggest that, in solution, VP6 trimers present a structural flexibility that is controlled by the presence of a zinc ion.
author Erk, Inge
Huet, Jean-Claude
Duarte, Mariela
Duquerroy, Stéphane
Rey, Felix
Cohen, Jean
Lepault, Jean
author_facet Erk, Inge
Huet, Jean-Claude
Duarte, Mariela
Duquerroy, Stéphane
Rey, Felix
Cohen, Jean
Lepault, Jean
author_sort Erk, Inge
title A Zinc Ion Controls Assembly and Stability of the Major Capsid Protein of Rotavirus
title_short A Zinc Ion Controls Assembly and Stability of the Major Capsid Protein of Rotavirus
title_full A Zinc Ion Controls Assembly and Stability of the Major Capsid Protein of Rotavirus
title_fullStr A Zinc Ion Controls Assembly and Stability of the Major Capsid Protein of Rotavirus
title_full_unstemmed A Zinc Ion Controls Assembly and Stability of the Major Capsid Protein of Rotavirus
title_sort zinc ion controls assembly and stability of the major capsid protein of rotavirus
publisher American Society for Microbiology
publisher_facet American Society for Microbiology
publishDate 2003
url https://ncbi.nlm.nih.gov/pmc/articles/PMC149495/
https://ncbi.nlm.nih.gov/pubmed/12610135
https://ncbi.nlm.nih.govhttp://dx.doi.org/10.1128/JVI.77.6.3595-3601.2003
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