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Related Experiment Videos

Actin-depolymerizing factor mediates Rac/Rop GTPase-regulated pollen tube growth.

Christine Y-h Chen1, Alice Y Cheung, Hen-ming Wu

  • 1Molecular and Cell Biology Program, University of Massachusetts, Amherst, Massachusetts 01003, USA.

The Plant Cell
|January 2, 2003
PubMed
Summary

Tobacco pollen actin-depolymerizing factors (ADFs)/cofilins are crucial for actin dynamics. Phosphorylation of NtADF1 by NtRac1 regulates its activity, controlling pollen tube growth.

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Area of Science:

  • Plant Biology
  • Cell Biology
  • Molecular Biology

Background:

  • Pollen tube elongation relies on a dynamic actin cytoskeleton.
  • Actin-depolymerizing factors (ADFs)/cofilins regulate actin dynamics by binding and severing actin filaments.
  • Phosphorylation of ADFs/cofilins by Rho GTPases is a key regulatory mechanism.

Purpose of the Study:

  • To investigate the role of NtADF1, a tobacco pollen-specific ADF, in pollen tube growth.
  • To determine if NtADF1 activity is regulated by phosphorylation.
  • To elucidate the interaction between NtADF1 and NtRac1 in regulating pollen tube actin cytoskeleton.

Main Methods:

  • Overexpression of NtADF1 and mutant forms (NtADF1(S6A), NtADF1(S6D)) in tobacco pollen.
  • Functional assay using NtRac1-induced defective pollen tube phenotype.

Related Experiment Videos

  • Assessment of actin binding activity of NtADF1 using green fluorescent protein (GFP) tagging.

    • Main Results:

    • Overexpression of NtADF1 suppressed the isotropic growth phenotype induced by NtRac1.
    • Mutant NtADF1(S6A) showed increased activity, while NtADF1(S6D) showed reduced activity in counteracting NtRac1 effects.
    • NtRac1 overexpression diminished GFP-NtADF1 actin binding but not GFP-NtADF1(S6A).
    • Constitutively active NtRac1 increased the ratio of phosphorylated to nonphosphorylated ADFs.

    Conclusions:

    • NtADF1 acts in a common pathway with NtRac1 to regulate pollen tube growth.
    • Phosphorylation at Ser-6 of NtADF1 is crucial for its integration into the NtRac1 signaling pathway.
    • NtRac1 modulates NtADF1 activity via phosphorylation at Ser-6, regulating actin dynamics during pollen tube growth.