A zinc ion controls assembly and stability of the major capsid protein of rotavirus
| Autor: | Jean Lepault, Jean-Claude Huet, Jean Cohen, Inge Erk, Stéphane Duquerroy, Félix A. Rey, Mariela Duarte |
|---|---|
| Přispěvatelé: | Virologie moléculaire et structurale (VMS), Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS), Biochimie bactérienne (BIOBAC), Institut National de la Recherche Agronomique (INRA), Commissariat à l'énergie atomique et aux énergies alternatives (CEA) |
| Jazyk: | angličtina |
| Rok vydání: | 2003 |
| Předmět: |
Gene Expression Regulation
Viral Rotavirus EXPRESSION Transcription Genetic medicine.medical_treatment viruses Immunology Mutant chemistry.chemical_element Trimer Zinc Spodoptera Biology Microbiology Serine 03 medical and health sciences Virology medicine Animals PARTICLES CORE Antigens Viral Cells Cultured Histidine 3-DIMENSIONAL STRUCTURE 030304 developmental biology Zinc finger 0303 health sciences Protease ATOMIC-STRUCTURE 030306 microbiology Structure and Assembly Virus Assembly virus diseases CRYOELECTRON MICROSCOPY Nucleopolyhedroviruses Microscopy Electron chemistry Capsid Biochemistry Insect Science Mutation [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology Capsid Proteins COMPLEXES |
| Zdroj: | Journal of Virology Journal of Virology, American Society for Microbiology, 2003, 77 (6), pp.3595-3601. ⟨10.1128/JVI.77.6.3595-3601.2003⟩ Journal of Virology, 2003, 77 (6), pp.3595-3601. ⟨10.1128/JVI.77.6.3595-3601.2003⟩ |
| ISSN: | 0022-538X 1098-5514 |
| Popis: | 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, 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. |
| Databáze: | OpenAIRE |
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