Abstract
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.
Competing Interest Statement
The authors have declared no competing interest.
Footnotes
Correct spelling of the author "Michal Jasinski".
Abbreviations
- IAA
- indole-3-acetic acid
- BA
- benzoic acid
- PAT
- polar auxin transport
- ABCB
- ATP-binding cassette protein subfamily B
- PGP
- P-glycoprotein
- TWD1
- Twisted Dwarf1
- DGT
- DIAGEOTROPICA
- MDR
- multi-drug resistance
- NPA
- 1-N-naphthylphthalamic acid
- FKBP
- FK506-binding protein
- WT
- wild-type
- NBD
- nucleotide binding domain
- TMD
- transmembrane domain
- ICL
- intracellular loop
- FKBD
- FK506-binding domain
- AUX1/LAX
- AUXIN1-RESISTANT1/LIKE
- PIN
- PIN-FORMED
- CFTR
- cystic fibrosis transmembrane conductance regulator
- PPIase
- cis-trans peptidyl-prolyl isomerase
- PM
- plasma membrane
- ER
- endoplasmic reticulum
- BA
- benzoic acid