Geisler, M. | Search bioRxiv

bioRxiv2020

ABCB-mediated auxin transport in outer root tissues regulates lateral root spacing in Arabidopsis

Root branching is an important strategy to explore efficiently large volumes of soil. To economize this process, lateral roots (LR) are formed along the growing root at discrete positions that are instructed by oscillating auxin signals derived from the lateral root cap (LRC). This assumes that auxin moves from the LRC across multiple layers to accumulate in the pericycle. Here, we identified, using gene silencing and CRISPR based approaches, a group of five genetically linked, closely related ABCBs that control LR spacing by modulating the amplitude of the auxin oscillation. The transporters localize to the plasma membrane and reveal significant auxin export activity. These ABCBs are mainly expressed in the LRC and epidermis where they contribute to auxin transport towards the root oscillation zone. Our findings highlight the importance of auxin transport in the outer tissues of the root meristem to regulate LR spacing.

plant biology↗

bioRxiv2020

A conserved D/E-P motif in the nucleotide binding domain of plant ABCB/PGP-type ABC transporters defines their auxin transport capacity

Auxin transport activity of ABCB1 was suggested to be regulated by physical interaction with the FKBP42/Twisted Dwarf1 (TWD1), a bona fide peptidylprolyl cis-trans isomerase (PPIase), but all attempts to demonstrate such a PPIase activity on TWD1 have failed so far. By using a structure-based approach we have identified a series of surface-exposed proline residues in the C-terminal nucleotide binding fold and linker of Arabidopsis ABCB1 that do not alter ABCB1 protein stability or location but its catalytic transport activity. P1.008 was uncovered as part of a conserved signature D/E-P motif that seems to be specific for Auxin-Transporting ABCBs, we now refer to as ATAs. Beside the proline, also mutation of the acidic moiety prior to the proline abolishes auxin transport activity by ABCB1. So far, all higher plant ABCBs for that auxin transport was safely proven carry this conserved motif underlining its diagnostic potential. Introduction of this D/E-P motif into malate importer, ABCB14, increases both its malate and its background auxin transport activity, suggesting that this motif has an impact on transport capacity. The D/E-P1.008 motif is also important for ABCB1-TWD1 interaction and activation of ABCB1-mediated auxin transport by TWD1, supporting a scenario in that TWD1 acts as an activator of ABCB1 transport activity by means of its PPIase. In summary, our data imply a dual function for TWD1 acting as an ABCB co-chaperone required for ABCB biogenesis and as a putative activator of ABCB-mediated auxin transport by cis-trans isomerization of peptidyl-prolyl bonds.

biochemistry↗