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Tuning a timing device that regulates lateral root development in rice

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Abstract

Peptidyl Prolyl Isomerases (PPIases) accelerate cis–trans isomerization of prolyl peptide bonds. In rice, the PPIase LRT2 is essential for lateral root initiation. LRT2 displays in vitro isomerization of a highly conserved W–P peptide bond (104W–P105) in the natural substrate OsIAA11. OsIAA11 is a transcription repressor that, in response to the plant hormone auxin, is targeted to ubiquitin-mediated proteasomal degradation via specific recognition of the cis isomer of its 104W–P105 peptide bond. OsIAA11 controls transcription of specific genes, including its own, that are required for lateral root development. This auxin-responsive negative feedback circuit governs patterning and development of lateral roots along the primary root. The ability to tune LRT2 activity via mutagenesis is crucial for understanding and modeling the role of this bimodal switch in the auxin circuit and lateral root development. We present characterization of the thermal stability and isomerization rates of several LRT2 mutants acting on the OsIAA11 substrate. The thermally stable mutants display activities lower than that of wild-type (WT) LRT2. These include binding diminished but catalytically active P125K, binding incompetent W128A, and binding capable but catalytically incompetent H133Q mutations. Additionally, LRT2 homologs hCypA from human, TaCypA from Triticum aestivum (wheat) and PPIB from E. coli were shown to have 110, 50 and 60% of WT LRT2 activity on the OsIAA11 substrate. These studies identify several thermally stable LRT2 mutants with altered activities that will be useful for establishing relationships between cis–trans isomerization, auxin circuit dynamics, and lateral root development in rice.

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Acknowledgements

This work is dedicated in honor of Dr. Dennis Torchia and his pioneering contributions to the field of protein NMR spectroscopy and to the training of multiple generations of NMR spectroscopists. As one of those trainees, LKN is grateful for the lasting and significant impact of Dr. Torchia’s mentorship on her journey in science. This work was supported by the National Science Foundation Grant MCB-1615350 and by the National Institutes of Health under the Ruth L. Kirschstein National Research Service Award (2T32GM008267) from the National Institute of General Medical Sciences.

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  1. Lucila Andrea Acevedo

    Present address: Department of Biochemistry and Biophysics and the Abramson Family Cancer Research Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA

  2. Lucila Andrea Acevedo and Nathan E. Korson contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular Biology & Genetics, Cornell University, Ithaca, NY, 14853, USA

    Lucila Andrea Acevedo, Nathan E. Korson, Justin M. Williams & Linda K. Nicholson

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  1. Lucila Andrea Acevedo
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Correspondence to Linda K. Nicholson.

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Acevedo, L.A., Korson, N.E., Williams, J.M. et al. Tuning a timing device that regulates lateral root development in rice. J Biomol NMR 73, 493–507 (2019). https://doi.org/10.1007/s10858-019-00258-0

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