Skip to main content

Rho GTPase Activity Analysis in Plant Cells

  • Protocol
  • First Online:
Book cover Plant Signalling Networks

Part of the book series: Methods in Molecular Biology ((MIMB,volume 876))

Abstract

Rho-family small GTPases are conserved molecular switches of signaling networks in eukaryotic cells, and regulate many cellular responses, such as cytoskeletal reorganization, gene expression, and polarized vesicular trafficking. To understand the functions of Rho GTPase, it is important to investigate how the activity of Rho GTPase is regulated. Plant Rho-like GTPases (ROPs) are known to be regulated by hormones and environmental cues. A rapid activation of ROPs by a stimulus implies a direct signaling role for the ROP GTPases. Here, I describe an ROP activity assay that allows the measurement of ROP GTPase activity that occurs within seconds upon treatment with a stimulus. This method has been successfully used to investigate auxin activation of ROPs in plant cells and will be generally useful for measuring changes in ROP activity with high time resolution (Xu et al., Cell 143:99–110, 2010).

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Yang Z (2002) Small GTPases: versatile signaling switches in plants. Plant Cell 14(Suppl):S375–388

    PubMed  CAS  Google Scholar 

  2. Gu Y, Wang Z, Yang Z (2004) ROP/RAC GTPase: an old new master regulator for plant signaling. Curr Opin Plant Biol 7:527–536

    Article  PubMed  CAS  Google Scholar 

  3. Nagawa S, Xu T, Yang Z (2010) RHO GTPase in plants: conservation and invention of regulators and effectors. Small GTPases 1:78–88

    Article  PubMed  Google Scholar 

  4. Wu G, Gu Y, Li S, Yang Z (2001) A genome-wide analysis of Arabidopsis Rop-interactive CRIB motif-containing proteins that act as Rop GTPase targets. Plant Cell 13:2841–2856

    Article  PubMed  CAS  Google Scholar 

  5. Fu Y, Li H, Yang Z (2002) The ROP2 GTPase controls the formation of cortical fine F-actin and the early phase of directional cell expansion during Arabidopsis organogenesis. Plant Cell 14:777–794

    Article  PubMed  CAS  Google Scholar 

  6. `Fu Y, Gu Y, Zheng Z, Wasteneys G, Yang Z (2005) Arabidopsis interdigitating cell growth requires two antagonistic pathways with opposing action on cell morphogenesis. Cell 120:687–700

    Article  PubMed  CAS  Google Scholar 

  7. Gu Y, Fu Y, Dowd P, Li S, Vernoud V, Gilroy S, Yang Z (2005) A Rho family GTPase controls actin dynamics and tip growth via two counteracting downstream pathways in pollen tubes. J Cell Biol 169:127–138

    Article  PubMed  CAS  Google Scholar 

  8. Fu Y, Xu T, Zhu L, Wen M, Yang Z (2009) A ROP GTPase signaling pathway controls cortical microtubule ordering and cell expansion in Arabidopsis. Curr Biol 19:1827–1832

    Article  PubMed  CAS  Google Scholar 

  9. Xu T, Wen M, Nagawa S, Fu Y, Chen JG, Wu MJ, Perrot-Rechenmann C, Friml J, Jones AM, Yang Z (2010) Cell surface- and rho GTPase-based auxin signaling controls cellular interdigitation in Arabidopsis. Cell 143:99–110

    Article  PubMed  CAS  Google Scholar 

  10. Baxter-Burrell A, Yang Z, Springer PS, Bailey-Serres J (2002) RopGAP4-dependent Rop GTPase rheostat control of Arabidopsis oxygen deprivation tolerance. Science 296:2026–2028

    Article  PubMed  CAS  Google Scholar 

  11. Tao LZ, Cheung AY, Wu HM (2002) Plant Rac-like GTPases are activated by auxin and mediate auxin-responsive gene expression. Plant Cell 14:2745–2760

    Article  PubMed  CAS  Google Scholar 

  12. Sheen J (2001) Signal transduction in maize and Arabidopsis mesophyll protoplasts. Plant Physiol 127:1466–1475

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Tongda Xu .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2011 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Xu, T. (2011). Rho GTPase Activity Analysis in Plant Cells. In: Wang, ZY., Yang, Z. (eds) Plant Signalling Networks. Methods in Molecular Biology, vol 876. Humana Press. https://doi.org/10.1007/978-1-61779-809-2_10

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-809-2_10

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-808-5

  • Online ISBN: 978-1-61779-809-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics