A comprehensive analysis of the 14-3-3 interactome in barley leaves using a complementary proteomics and two-hybrid approach.

Content Provider	Semantic Scholar
Author	Schoonheim, Peter J. Veiga, Helena Pereira, Daniel Da Costa Friso, Giulia Wijk, Klaas J. Van Boer, Albertus H. De
Copyright Year	2007
Abstract	This study describes the identification of over 150 target proteins of the five 14-3-3 isoforms in 7-d-old barley (Hordeum vulgare) cv Himalaya seedlings using yeast two-hybrid screens complemented with 14-3-3 protein affinity purification and tandem mass spectrometry. Independent experiments for a subset of genes confirmed the yeast two-hybrid interactions, demonstrating a low false positive identification rate. These combined approaches resulted in the identification of more than 150 putative targets; 15% were previously reported to be 14-3-3 interactors, including, for example, Serpin, RF2A, WPK4 kinase, P-type proton-translocating adenosine triphosphatase, EF1A, glutamine synthetase, and invertases. The affinity purification resulted in 30 interactors, of which 44% function in metabolism, while the yeast two-hybrid screens identified 132 different proteins, with 35% of the proteins involved in signal transduction. A number of proteins have a well-described function in hormonal signaling, such as the auxin transport protein PIN1 and NPH3 and components of the brassinosteroid pathway, such as the receptor kinase BAK1 (OsPERK1) and BRI1-kinase domain-interacting protein 129. However, 14-3-3 interactions with these signal mediators have not been confirmed in the affinity purification. Confirmations of the 14-3-3 interaction with the three ABF-like transcription factors are shown using far western analysis. Also, a REPRESSION OF SHOOT GROWTH ortholog named RF2A was identified; these transcription factors play important roles in the abscisic acid and gibberellin pathways, respectively. We speculate that 14-3-3 proteins have a role in cross talk between these hormonal pathways. The specificity and complementary nature of both the affinity purification and the yeast two-hybrid approaches is discussed.
File Format	PDF HTM / HTML
DOI	10.1104/pp.106.090159
PubMed reference number	17172288
Journal	Medline
Volume Number	143
Issue Number	2
Alternate Webpage(s)	http://dare.ubvu.vu.nl/bitstream/handle/1871/20582/PMC1803744.pdf?isAllowed=y&sequence=1
Alternate Webpage(s)	http://www.plantphysiol.org/content/plantphysiol/143/2/670.full.pdf
Alternate Webpage(s)	https://doi.org/10.1104/pp.106.090159
Journal	Plant physiology
Language	English
Access Restriction	Open
Content Type	Text
Resource Type	Article