title: Untethering the TIR1 auxin receptor from the SCF complex increases its stability and inhibits auxin response.
abstract: Plant genomes encode large numbers of F-box proteins (FBPs), the substrate recognition subunit of SKP1-CULLIN-F-box (SCF) ubiquitin ligases. There are ~700 FBPs inArabidopsis, most of which are uncharacterized. TIR1 is among the best-studied plant FBPs and functions as a receptor for the plant hormone auxin. Here we use a yeast two-hybrid system to identify novel TIR1 mutants with altered properties. The analysis of these mutants reveals that TIR1 associates with the CULLIN1 (CUL1) subunit of the SCF through the N-terminal H1 helix of the F-box domain. Mutations that untether TIR1 from CUL1 stabilize the FBP and cause auxin resistance and associated growth defects, probably by protecting TIR1 substrates from degradation. Based on these results we propose that TIR1 is subject to autocatalytic degradation when assembled into an SCF. Further, our results suggest a general method for determining the physiological function of uncharacterized FBPs. Finally, we show that a key amino acid variation in the F-box domain of auxin signalling F-box (AFB1), a closely related FBP, reduces its ability to form an SCF, resulting in an increase in AFB1 levels.
authors.affiliation: Howard Hughes Medical Institute and Section of Cell and Developmental Biology, University of California San Diego, La Jolla, California 92093, USA.
authors.forename: Hong
authors.initials: H
authors.lastname: Yu
authors.affiliation: Howard Hughes Medical Institute and Section of Cell and Developmental Biology, University of California San Diego, La Jolla, California 92093, USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Biology, University of Washington, Seattle, Washington 98195, USA.
authors.forename: Britney L
authors.initials: BL
authors.lastname: Moss
authors.affiliation: Howard Hughes Medical Institute and Section of Cell and Developmental Biology, University of California San Diego, La Jolla, California 92093, USA.
authors.forename: Bastiaan O R
authors.initials: BO
authors.lastname: Bargmann
authors.affiliation: Howard Hughes Medical Institute and Section of Cell and Developmental Biology, University of California San Diego, La Jolla, California 92093, USA.
authors.forename: Renhou
authors.initials: R
authors.lastname: Wang
authors.affiliation: Howard Hughes Medical Institute and Section of Cell and Developmental Biology, University of California San Diego, La Jolla, California 92093, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Prigge
authors.affiliation: Department of Biology, University of Washington, Seattle, Washington 98195, USA.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Nemhauser
authors.affiliation: Howard Hughes Medical Institute and Section of Cell and Developmental Biology, University of California San Diego, La Jolla, California 92093, USA.
authors.forename: Mark
authors.initials: M
authors.lastname: Estelle