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Localization of Rac2 via the C terminus and aspartic acid 150 specifies superoxide generation, actin polarity and chemotaxis in neutrophils

Nature Immunology volume 5, pages 744–751 (2004)Cite this article

Abstract

Despite having a high degree of sequence similarity, the Rho guanosine triphosphatases Rac1 and Rac2 regulate distinct functions in neutrophils. Here we demonstrate that the unique Rac2 localization and functions in neutrophils are regulated by two separate C-terminal motifs, the hypervariable domain and aspartic acid 150, one of which has not previously been linked to the function of Rho GTPases. In addition, we show an unexpected dependence of Rac1 localization on Rac2 activity in these same cells, demonstrating a degree of crosstalk between two closely related Rho GTPases. Thus, we have defined specific sequences in Rac that specify subcellular localization and determine the specificity of Rac2 in neutrophil chemotaxis and superoxide generation.

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Figure 1: Rac2 has distinct functions compared with Rac1 in primary neutrophils.
Figure 2: Rac1 and Rac2 show distinct cellular localization.
Figure 3: The RQQKRP motif determines the specificity of Rac2 in superoxide production but not directed migration.
Figure 4: Amino acid D150 and the RQQKRP motif determine the specificity of Rac2 in chemotaxis.
Figure 5: Amino acid D150 and the RQQKRP motif of Rac2 regulate polarized fMLP-induced F-actin assembly.
Figure 6: The specificity of Rac2 versus Rac1 is correlated with a specific subcellular distribution.
Figure 7: Polarized distribution of Rac2 is necessary for pseudopodia formation.

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Acknowledgements

We thank J. Robbins and H. Osinska for help with confocal analysis; D. Marmer and the Flow Cytometry Core at Cincinnati Children's Hospital Medical Center for assistance with cell sorting; S. Homan and V. Summey-Harner for animal husbandry; and S. Wheeler and K. Steward for administrative assistance. Supported by National Institutes of Health (RO1 DK62757 to D.A.W.).

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Authors and Affiliations

  1. Division of Experimental Hematology, Cincinnati Children's Hospital Research Foundation, University of Cincinnati College of Medicine, Cincinnati, 45229, Ohio, USA

    Marie-Dominique Filippi, Chad E Harris, Yi Gu, Yi Zheng & David A Williams

  2. Division of Pediatric Informatics, Cincinnati Children's Hospital Research Foundation, University of Cincinnati College of Medicine, Cincinnati, 45229, Ohio, USA

    Jarek Meller

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  1. Marie-Dominique Filippi
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Correspondence to David A Williams.

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Supplementary information

Supplementary Fig. 1

Confocal microscopy analysis of Rac mutant localization 15 sec after fMLP stimulation as compared with plasma membrane localization. (PDF 1401 kb)

Supplementary Table 1

Specificity of Rac2 functions is correlated with its subcellular distribution. (PDF 16 kb)

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Filippi, MD., Harris, C., Meller, J. et al. Localization of Rac2 via the C terminus and aspartic acid 150 specifies superoxide generation, actin polarity and chemotaxis in neutrophils. Nat Immunol 5, 744–751 (2004). https://doi.org/10.1038/ni1081

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