Bimolecular fluorescence complementation and interaction of various Arabidopsis major intrinsic proteins expressed in yeast

TY - JOUR

T1 - Bimolecular fluorescence complementation and interaction of various Arabidopsis major intrinsic proteins expressed in yeast

AU - Murozuka, Emiko

AU - Hanisch, Susanne

AU - Günther-Pomorski, Thomas

AU - Jahn, Thomas Paul

AU - Schjørring, Jan Kofod

N1 - Special Issue: Functional Genomics

PY - 2013/7

Y1 - 2013/7

N2 - Tonoplast intrinsic proteins (TIPs) and plasma membrane intrinsic proteins (PIPs) form subgroups of plant major intrinsic proteins (MIPs) that channel water as well as various small neutral molecules across the tonoplast and plasma membrane. Most MIPs are believed to form homotetramers, while some plant PIPs have been shown to form heterotetramers composed of different isoforms. This study investigated in vivo molecular interactions between different Arabidopsis TIP isoforms and between TIPs and a PIP member. The interactions were assayed by bimolecular fluorescence complementation optimized for use in Saccharomyces cerevisiae as a heterologous expression system. Fluorescence of re-assembled Venus yellow fluorescent protein was monitored by fluorescence microscopy and flow cytometry. The results showed strong interactions between TIP1;2, TIP2;1 and TIP3;1. Surprisingly, the three TIP isoforms also interacted with PIP2;1. The potassium channel AKT1 was used as a negative control and exhibited no interaction with any of the MIPs. The observed interactions may play a role in targeting and regulation of MIPs in plants.

AB - Tonoplast intrinsic proteins (TIPs) and plasma membrane intrinsic proteins (PIPs) form subgroups of plant major intrinsic proteins (MIPs) that channel water as well as various small neutral molecules across the tonoplast and plasma membrane. Most MIPs are believed to form homotetramers, while some plant PIPs have been shown to form heterotetramers composed of different isoforms. This study investigated in vivo molecular interactions between different Arabidopsis TIP isoforms and between TIPs and a PIP member. The interactions were assayed by bimolecular fluorescence complementation optimized for use in Saccharomyces cerevisiae as a heterologous expression system. Fluorescence of re-assembled Venus yellow fluorescent protein was monitored by fluorescence microscopy and flow cytometry. The results showed strong interactions between TIP1;2, TIP2;1 and TIP3;1. Surprisingly, the three TIP isoforms also interacted with PIP2;1. The potassium channel AKT1 was used as a negative control and exhibited no interaction with any of the MIPs. The observed interactions may play a role in targeting and regulation of MIPs in plants.

U2 - 10.1111/ppl.12000

DO - 10.1111/ppl.12000

M3 - Journal article

C2 - 23163742

SN - 0031-9317

VL - 148

SP - 422

EP - 431

JO - Physiologia Plantarum

JF - Physiologia Plantarum

IS - 3

ER -