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WRKY23 is a component of the transcriptional network mediating auxin feedback on PIN polarity
(2018)
Auxin is unique among plant hormones due to its directional transport that is mediated by the polarly distributed PIN auxin transporters at the plasma membrane. The canalization hypothesis proposes that the auxin feedback on its polar flow is a crucial, plant-specific mechanism mediating multiple self-organizing developmental processes. Here, we used the auxin effect on the PIN polar localization in Arabidopsis thaliana roots as a proxy for the auxin feedback on the PIN polarity during canalization. We performed microarray experiments to find regulators of this process that act downstream of auxin. We identified genes that were transcriptionally regulated by auxin in an AXR3/IAA17-and ARF7/ARF19-dependent manner. Besides the known components of the PIN polarity, such as PID and PIP5K kinases, a number of potential new regulators were detected, among which the WRKY23 transcription factor, which was characterized in more detail. Gain-and loss-of-function mutants confirmed a role for WRKY23 in mediating the auxin effect on the PIN polarity. Accordingly, processes requiring auxin-mediated PIN polarity rearrangements, such as vascular tissue development during leaf venation, showed a higher WRKY23 expression and required the WRKY23 activity. Our results provide initial insights into the auxin transcriptional network acting upstream of PIN polarization and, potentially, canalization-mediated plant development.
The cytoskeletal motor protein kinesin-1 (conventional kinesin) is the fast carrier for intracellular cargo transport along microtubules. So far most studies aimed at investigating the transport properties of individual motor molecules. However, the transport in cells usually involves the collective work of more than one motor. In the present work, we have studied the movement of beads as artificial loads/organelles pulled by several kinesin-1 motors in vitro. For a wide range of motor coverage of the beads and different bead (cargo) sizes the transport parameters walking distance or run length, velocity and force generation are measured. The results indicate that the transport parameters are influenced by the number of motors carrying the bead. While the transport velocity slightly decreases, an increase in the run length was measured and higher forces are determined, when more motors are involved. The effective number of motors pulling a bead is estimated by measuring the change in the hydrodynamic diameter of kinesin-coated beads using dynamic light scattering. The geometrical constraints imposed by the transport system have been taken into account. Thus, results for beads of different size and motor-surface coverage could be compared. In addition, run length-distributions obtained for the smallest bead size were matched to theoretically calculated distributions. The latter yielded an average number of pulling motors, which is in agreement with the effective motor numbers determined experimentally.