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
Publication statusPublished - 2004 Aug
Externally publishedYes

Fingerprint

protein isoforms
Tobacco mosaic virus
genes
proteins
elicitors
recombinant proteins
Northern blotting
bacteriophages
Nicotiana tabacum
tobacco
sequence analysis
reverse transcriptase polymerase chain reaction
clones
screening
nucleotide sequences
phylogeny
assays

Keywords

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

ASJC Scopus subject areas

  • Plant Science

Cite this

Members of 14-3-3 protein isoforms interacting with the resistance gene product N and the elicitor of Tobacco mosaic virus. / Konagaya, Ken Ichi; Matsushita, Yasuhiko; Kasahara, Masahiro; Nyunoya, Hiroshi.

In: Journal of General Plant Pathology, Vol. 70, No. 4, 08.2004, p. 221-231.

Research output: Contribution to journalArticle

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