An Auxin-based Degron System for the Rapid Depletion of Proteins in Nonplant Cells

Overview

Journal Nat Methods

Specialties Biomedical Engineering
Pathology

Date 2009 Nov 17

PMID 19915560

Citations 896

Authors

Kohei Nishimura

Tatsuo Fukagawa

Haruhiko Takisawa

Tatsuo Kakimoto

Masato Kanemaki

Affiliations

Soon will be listed here.

Abstract

Plants have evolved a unique system in which the plant hormone auxin directly induces rapid degradation of the AUX/IAA family of transcription repressors by a specific form of the SCF E3 ubiquitin ligase. Other eukaryotes lack the auxin response but share the SCF degradation pathway, allowing us to transplant the auxin-inducible degron (AID) system into nonplant cells and use a small molecule to conditionally control protein stability. The AID system allowed rapid and reversible degradation of target proteins in response to auxin and enabled us to generate efficient conditional mutants of essential proteins in yeast as well as cell lines derived from chicken, mouse, hamster, monkey and human cells, thus offering a powerful tool to control protein expression and study protein function.

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References

Schulman B, Carrano A, Jeffrey P, Bowen Z, Kinnucan E, Finnin M . Insights into SCF ubiquitin ligases from the structure of the Skp1-Skp2 complex. Nature. 2000; 408(6810):381-6. DOI: 10.1038/35042620. View

Folkes L, Dennis M, Stratford M, Candeias L, Wardman P . Peroxidase-catalyzed effects of indole-3-acetic acid and analogues on lipid membranes, DNA, and mammalian cells in vitro. Biochem Pharmacol. 1999; 57(4):375-82. DOI: 10.1016/s0006-2952(98)00323-2. View

Zhang J, Zheng N, Zhou P . Exploring the functional complexity of cellular proteins by protein knockout. Proc Natl Acad Sci U S A. 2003; 100(24):14127-32. PMC: 283557. DOI: 10.1073/pnas.2233012100. View

Picard D . Posttranslational regulation of proteins by fusions to steroid-binding domains. Methods Enzymol. 2000; 327:385-401. DOI: 10.1016/s0076-6879(00)27291-1. View

Fukagawa T, Mikami Y, Nishihashi A, Regnier V, Haraguchi T, Hiraoka Y . CENP-H, a constitutive centromere component, is required for centromere targeting of CENP-C in vertebrate cells. EMBO J. 2001; 20(16):4603-17. PMC: 125570. DOI: 10.1093/emboj/20.16.4603. View

Teale W, Paponov I, Palme K . Auxin in action: signalling, transport and the control of plant growth and development. Nat Rev Mol Cell Biol. 2006; 7(11):847-59. DOI: 10.1038/nrm2020. View

Tan X, Calderon-Villalobos L, Sharon M, Zheng C, Robinson C, Estelle M . Mechanism of auxin perception by the TIR1 ubiquitin ligase. Nature. 2007; 446(7136):640-5. DOI: 10.1038/nature05731. View

Armstrong C, Goldberg D . An FKBP destabilization domain modulates protein levels in Plasmodium falciparum. Nat Methods. 2007; 4(12):1007-9. DOI: 10.1038/nmeth1132. View

Zhou P . Targeted protein degradation. Curr Opin Chem Biol. 2005; 9(1):51-5. DOI: 10.1016/j.cbpa.2004.10.012. View

10.

Haruki H, Nishikawa J, Laemmli U . The anchor-away technique: rapid, conditional establishment of yeast mutant phenotypes. Mol Cell. 2008; 31(6):925-32. DOI: 10.1016/j.molcel.2008.07.020. View

11.

Schneekloth Jr J, Fonseca F, Koldobskiy M, Mandal A, Deshaies R, Sakamoto K . Chemical genetic control of protein levels: selective in vivo targeted degradation. J Am Chem Soc. 2004; 126(12):3748-54. DOI: 10.1021/ja039025z. View

12.

Sauer B, Henderson N . Site-specific DNA recombination in mammalian cells by the Cre recombinase of bacteriophage P1. Proc Natl Acad Sci U S A. 1988; 85(14):5166-70. PMC: 281709. DOI: 10.1073/pnas.85.14.5166. View

13.

Elbashir S, Harborth J, Lendeckel W, Yalcin A, Weber K, Tuschl T . Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature. 2001; 411(6836):494-8. DOI: 10.1038/35078107. View

14.

Su X, Bernal J, Venkitaraman A . Cell-cycle coordination between DNA replication and recombination revealed by a vertebrate N-end rule degron-Rad51. Nat Struct Mol Biol. 2008; 15(10):1049-58. DOI: 10.1038/nsmb.1490. View

15.

de Melo M, de Lima T, Pithon-Curi T, Curi R . The mechanism of indole acetic acid cytotoxicity. Toxicol Lett. 2004; 148(1-2):103-11. DOI: 10.1016/j.toxlet.2003.12.067. View

16.

Labib K, Tercero J, Diffley J . Uninterrupted MCM2-7 function required for DNA replication fork progression. Science. 2000; 288(5471):1643-7. DOI: 10.1126/science.288.5471.1643. View

17.

Banaszynski L, Chen L, Maynard-Smith L, Ooi A, Wandless T . A rapid, reversible, and tunable method to regulate protein function in living cells using synthetic small molecules. Cell. 2006; 126(5):995-1004. PMC: 3290523. DOI: 10.1016/j.cell.2006.07.025. View

18.

Ruegger M, Dewey E, Gray W, Hobbie L, Turner J, Estelle M . The TIR1 protein of Arabidopsis functions in auxin response and is related to human SKP2 and yeast grr1p. Genes Dev. 1998; 12(2):198-207. PMC: 316440. DOI: 10.1101/gad.12.2.198. View

19.

Gossen M, Bujard H . Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc Natl Acad Sci U S A. 1992; 89(12):5547-51. PMC: 49329. DOI: 10.1073/pnas.89.12.5547. View

20.

Herm-Gotz A, Agop-Nersesian C, Munter S, Grimley J, Wandless T, Frischknecht F . Rapid control of protein level in the apicomplexan Toxoplasma gondii. Nat Methods. 2007; 4(12):1003-5. PMC: 2601725. DOI: 10.1038/nmeth1134. View