Two‐hybrid protein–protein interaction analysis in Arabidopsis protoplasts: establishment of a heterodimerization map of group C and group S bZIP transcription factors

Two‐hybrid protein–protein interaction analysis in Arabidopsis protoplasts: establishment of... Summary In vivo protein–protein interactions are frequently studied by means of yeast two‐hybrid analysis. However, interactions detected in yeast might differ considerably in the plant system. Based on GAL4 DNA‐binding (BD) and activation domains (AD) we established an Arabidopsis protoplast two‐hybrid (P2H) system. The use of Gateway®‐compatible vectors enables the high‐throughput screening of protein–protein interactions in plant cells. The efficiency of the system was tested by examining the homo‐ and heterodimerization properties of basic leucine zipper (bZIP) transcription factors. A comprehensive heterodimerization matrix of Arabidopsis thaliana group C and group S bZIP transcription factors was generated by comparing the results of yeast and protoplast two‐hybrid experiments. Surprisingly, almost no homodimerization but rather specific and selective heterodimerization was detected. Heterodimers were preferentially formed between group C members (AtbZIP9, ‐10, ‐25, ‐63) and members of group S1 (AtbZIP1, ‐2, ‐11, ‐44, ‐53). In addition, significant but low‐affinity interactions were detected inside group S1, S2 or C AtbZIPs, respectively. As a quantitative approach, P2H identified weak heterodimerization events which were not detected in the yeast system. Thus, in addition to cell biological techniques, P2H is a valuable tool for studying protein–protein interaction in living plant cells. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png The Plant Journal Wiley

Two‐hybrid protein–protein interaction analysis in Arabidopsis protoplasts: establishment of a heterodimerization map of group C and group S bZIP transcription factors

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Publisher
Wiley
Copyright
Copyright © 2006 Wiley Subscription Services, Inc., A Wiley Company
ISSN
0960-7412
eISSN
1365-313X
D.O.I.
10.1111/j.1365-313X.2006.02731.x
Publisher site
See Article on Publisher Site

Abstract

Summary In vivo protein–protein interactions are frequently studied by means of yeast two‐hybrid analysis. However, interactions detected in yeast might differ considerably in the plant system. Based on GAL4 DNA‐binding (BD) and activation domains (AD) we established an Arabidopsis protoplast two‐hybrid (P2H) system. The use of Gateway®‐compatible vectors enables the high‐throughput screening of protein–protein interactions in plant cells. The efficiency of the system was tested by examining the homo‐ and heterodimerization properties of basic leucine zipper (bZIP) transcription factors. A comprehensive heterodimerization matrix of Arabidopsis thaliana group C and group S bZIP transcription factors was generated by comparing the results of yeast and protoplast two‐hybrid experiments. Surprisingly, almost no homodimerization but rather specific and selective heterodimerization was detected. Heterodimers were preferentially formed between group C members (AtbZIP9, ‐10, ‐25, ‐63) and members of group S1 (AtbZIP1, ‐2, ‐11, ‐44, ‐53). In addition, significant but low‐affinity interactions were detected inside group S1, S2 or C AtbZIPs, respectively. As a quantitative approach, P2H identified weak heterodimerization events which were not detected in the yeast system. Thus, in addition to cell biological techniques, P2H is a valuable tool for studying protein–protein interaction in living plant cells.

Journal

The Plant JournalWiley

Published: Jun 1, 2006

References

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