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In vitro observation of the molecular interaction between NodD and its inducer naringenin as monitored by fluorescence resonance energy transfer.

Authors
  • Li, Fengqing
  • Hou, Bihe
  • Chen, Lei
  • Yao, Zhujun
  • Hong, Guofan
Type
Published Article
Journal
Acta Biochimica et Biophysica Sinica
Publisher
Oxford University Press
Publication Date
Sep 01, 2008
Volume
40
Issue
9
Pages
783–789
Identifiers
PMID: 18776990
Source
Medline
License
Unknown

Abstract

At initial stages in the Rhizobium legume symbiosis, most nodulation genes are controlled by NodD protein and plant inducers. Some genetic studies and other reports have suggested that NodD may be activated by its direct interaction with plant inducers. However, there has been no molecular evidence of such an inducing interaction. In this paper, we used fluorescence resonance energy transfer technique to see whether such an interaction exists between NodD and its activator, naringenin, in vitro. The tetracysteine motif (Cys-Cys-Pro-Gly-Cys-Cys) was genetically inserted into NodD to label NodD with 4',5'-bis(1,3,2-dithioarsolan-2-yl) fluorescein (FlAsH). Naringenin was labeled with fluorescein by chemical linking. In the fluorescence resonance energy transfer experiments in vitro, the fluorescence intensity of one acceptor, NodD(90R6)-FlAsH, increased by 13%. This suggests that NodD may directly interact with inducer naringenin in vitro and that the reaction centre is likely near hinge region 1 of NodD.

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