Journal
AMINO ACIDS
Volume 40, Issue 2, Pages 381-392Publisher
SPRINGER WIEN
DOI: 10.1007/s00726-010-0643-y
Keywords
Xanthomonas oryzae; Hpa1; Coiled-coil; Oligomerization; alpha-helix; Hypersensitive response
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Funding
- National Key Basic Research Plan of China [2003CB114204, 2006CB101902]
- National Key project of China [2004BA901A36]
- Key Project of Science and Technology of Jiangsu [BE-2005-604]
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Hpa1(Xoo) (harpin) is a type III secreted protein of the rice blight bacterial pathogen Xanthomonas oryzae pv. oryzae that elicits a hypersensitive response (HR) in nonhost tobacco. Hpa1(Xoo) is predicted to contain two potential coiled-coil (CC) regions, one at the N-terminus with a high probability of formation, and one at the C-terminus with a lower probability of formation. We constructed several CC-equivalent peptides by a chemosynthetic method, and investigated the structure-function of the predicted Hpa1(Xoo) CC regions, using biophysical and biochemical approaches. Both peptides elicited an HR in tobacco. Mutant versions of the N- and C-terminal peptides that were predicted to disrupt or favor CC formation were generated. The resulting altered HR activity and oligomerization indicated that the N-terminal CC region is essential for eliciting HR, but the C-terminus is not. The results also indicate that a 14-residue fragment (LDQLLCQLISALLQ) within the N-terminal CC region is a minimal and independent functional element for HR-induction in tobacco leaves. We propose that HR-induction requires a specific oligomerization of the CC regions of Hpa1(Xoo).
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