Protein–protein interactions of tandem affinity purification-tagged protein kinases in rice

Authors

Jai S. Rohila, University of Nebraska - Lincoln
Mei Chen, University of Nebraska - LincolnFollow
Shuo Chen, University of Nebraska - LincolnFollow
Johann Chen, University of California, Davis
Ronald CernyFollow
Chris Dardick, University of California, Davis
Patrick Canlas, University of California, DavisFollow
Xia Xu, University of California, DavisFollow
Michael Gribskov, Purdue UniversityFollow
Siddhartha KanrarFollow
Jian-Kang Zhu, University of California, Riverside
P C. RonaldFollow
Michael E. Fromm, University of Nebraska - LincolnFollow

Date of this Version

April 2006

Comments

Published in The Plant Journal 46:1 (2006), pp.1–13. doi: 10.1111/j.1365-313X.2006.02671.x Copyright © 2006 Jai S. Rohila, Mei Chen, Shuo Chen, Johann Chen, Ronald Cerny, Chris Dardick, Patrick Canlas, Xia Xu, Michael Gribskov, Siddhartha Kanrar, Jian-Kang Zhu, Pamela Ronald, and Michael E. Fromm. Published by Blackwell Publishing Ltd.

Abstract

Forty-one rice cDNAs encoding protein kinases were fused to the tandem affinity purification (TAP) tag and expressed in transgenic rice plants. The TAP-tagged kinases and interacting proteins were purified from the T1 progeny of the transgenic rice plants and identified by mass spectrometry. Ninety-five percent of the TAP-tagged kinases were recovered. Fifty-six percent of the TAP-tagged kinases were found to interact with other rice proteins. A number of these interactions were consistent with known protein complexes found in other species, validating the TAP-tag method in rice plants. Phosphorylation sites were identified on four of the kinases that interacted with either 14-3-3 proteins or cyclins.