Localization of domains within the Drosophila Ref(2)P protein involved in the intracellular control of sigma rhabdovirus multiplication.
The ref(2)P gene of Drosophila melanogaster interferes with sigma rhabdovirus multiplication. This gene is highly variable, and the different alleles are considered permissive or restrictive according to their effects on virus replication. In all cases, the mechanisms involve intracellular interactions between the sigma virus and Ref(2)P proteins. We showed that the N-terminal domain of the Ref(2)P protein was required for its activity in vivo. The protein was inactive in the null p(od)2 mutant when its first 82 amino acids were deleted. The p delta n gene was constructed so that the first 91 amino acids coded for by the restrictive alleles could be expressed in vivo. It was active in a transformed line. This sequence was sufficient to impart a restrictive phenotype to an adult D. melanogaster fly after it was injected with the virus. However, the truncated protein expressed by p delta n did not have an effect on the hereditary transmission of the sigma virus to the offspring of the infected flies, even though it contained the restriction site. The native Ref(2)P protein has been previously shown to have conformation-dependent epitopes common with some of those of the viral N protein. We demonstrated the following. (i) These epitopes were found in a domain of the Ref(2)P protein distinct from the site involved in restriction. (ii) They were modified in the N protein of the haP7 sigma virus mutant selected as being adapted to the restrictive alleles of the ref(2)P gene; only one mutation in the N gene, leading to an amino acid substitution, distinguished the haP7 mutant from the original virus. (iii) The virus strains partially or totally adapted to the effects of the full restrictive protein expressed by pp were always found to multiply to a lesser extent in the presence of the protein expressed by p delta n. These data suggest that two distinct domains of the Ref(2)P protein are involved in the control of sigma virus multiplication.
Wyers
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Centre de Génétique Moleculaire, Centre National de la Recherche Scientifique, Gif sur Yvette, France.
Petitjean
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AM
Dru
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Gay
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Contamine
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D000483:Alleles; D000818:Animals; D001483:Base Sequence; D002213:Capsid / Q000276:immunology / Q000502:physiology; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000821:virology*; D018604:Epitope Mapping; D015870:Gene Expression; D008969:Molecular Sequence Data; D009154:Mutation; D009687:Nuclear Proteins; D011506:Proteins / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D012209:Rhabdoviridae / Q000502:physiology*; D013329:Structure-Activity Relationship; D014758:Viral Core Proteins / Q000276:immunology / Q000502:physiology
D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D009687:Nuclear Proteins; D011506:Proteins; D014758:Viral Core Proteins; C080596:ref(2)P protein, Drosophila