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

Abstract

SummaryForty-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 TAPtagged 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.

Genetic and physical fine mapping of the novel brown midrib gene bm6 in maize (Zea mays L.) to a 180 kb region on chromosome 2

“…In rice, a calmodulin-binding motif protein was reported to interact with wallassociated kinase (WAK)-2 like protein (Rohila et al 2006). WAKs are receptor like protein kinase, which are tightly linked with cell wall (He et al 1996), and are involved in regulation of cell elongation (Lally et al 2001) and expansion (Wagner and Kohorn 2001).…”

“…GRMZ2G 062396 is homologous to the naringenin 2-oxoglutarate 3-dioxygenasease encoding gene (F3H). The rice homologous counterpart of GRMZM2G348909 is an IQ calmodulinbinding motif family gene, which might interact with cell wall related kinase WAK2 (Rohila et al 2006).…”

Transferring an optimized TAP-toolbox for the isolation of protein complexes to a portfolio of rice tissues

“…CDKD was previously shown to form a heterotrimeric CDK-activating kinase (CAK) complex with a regulatory CYCLIN H (CYCH) subunit and the assembly factor ''MENAGE A TROIS 1'' (MAT1) both in rice (Rohila et al 2006) and Arabidopsis (Van Leene et al 2010). In the same studies, the other sub-complex of the general transcription factor II H (TFIIH), i.e.…”

“…A last, more highthroughput effort was presented by Rohila and co-workers. They identified interaction partners for 23 % of the 129 rice kinases screened, starting from the shoots of 6-8-week-old seedlings (Rohila et al 2006(Rohila et al , 2009). The rather low success rate in this study emphasizes that creating a comprehensive picture of possible interactions for a given complex in plants is a daunting task and that there is room for improvement of TAP protocols for plants.…”

“…We assessed the performance of the GS tag in rice callus, by comparing the interaction datasets resulting from purifications of the rice CYCLIN-DEPENDENT KINA-SES REGULATORY SUBUNIT 1 (CKS1) with the previously used 'improved TAP tag' or 'TAPi' tag (Rohila et al 2006). Then we applied the GS tag-based procedure to screen interaction partners for the rice ANAPHASE PROMOTING COMPLEX SUBUNIT 10 (APC10) and CYCLIN-DEPENDENT KINASE D (CDKD) proteins in a variety of rice tissues, including dissected organ parts.…”