Members of 14-3-3 protein isoforms interacting with the resistance gene product N and the elicitor of Tobacco mosaic virus

Ken Ichi Konagaya, Yasuhiko Matsushita, Masahiro Kasahara, Hiroshi Nyunoya

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

A functional cDNA for the resistance gene N from Nicotiana tabacum cv. Samsun NN to Tobacco mosaic virus (TMV) was obtained by reverse transcription polymerase chain reaction. Recombinant proteins containing various domains of the N protein were used in far-Western screening for the N-interacting protein from tobacco cDNA expression libraries. Nucleotide sequence analysis revealed that 199 cDNAs of 217 positive clones screened from 1.6 × 106 phage plaques were related to the 14-3-3 gene family. Phylogenetic analysis indicated that the 14-3-3 proteins were divided into 17 different 14-3-3 isoforms, including hitherto unidentified novel isoforms i-1 and i-2. In vitro binding assays showed that the probes TIR and LRR domains of N protein and helicase (HEL) domain of TMV replicase bound significantly to 14-3-3 isoforms of groups ω and ψ. These domains barely bound to those of group κ and did not bind to that of group ε. The same probes did not bind to any domains of N protein or the viral HEL domain. Northern blotting showed an increase in 14-3-3 isoform h transcripts coinciding with PR-1a induction after TMV inoculation. The 14-3-3 isoform h as a probe had significant binding to the LRR domain of N protein and the viral HEL domain, but there was only minor binding to the TIR domain. These results raise the possibility that 14-3-3 acts as a scaffolding or adaptor between the N protein and the helicase domain to support the interaction between the R gene product and the viral elicitor.

Original languageEnglish
Pages (from-to)221-231
Number of pages11
JournalJournal of General Plant Pathology
Volume70
Issue number4
DOIs
Publication statusPublished - 2004 Aug

Keywords

  • 14-3-3
  • Helicase
  • N gene
  • Nicotiana tabacum
  • Resistance
  • Tobacco mosaic virus

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Plant Science

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