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Neutrophil direction sensing and superoxide production linked by the GTPase-activating protein GIT2

Nature Immunology volume 7, pages 724–731 (2006)Cite this article

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Abstract

In neutrophils, superoxide anion production generally accompanies chemotaxis and functions in killing invading pathogens. The GIT2 GTPase-activating protein binds to the guanine nucleotide–exchange factor αPIX. Here we show that GIT2 was necessary for directional chemotaxis and for the suppression of superoxide production in G protein–coupled receptor–stimulated neutrophils. GIT2 was also necessary for the orientation of superoxide production toward chemoattractant sources. GIT2 suppressed the activity of ADP ribosylation factor 1 and was a component of the Gβγ subunit–mediated direction-sensing machinery 'downstream' of G protein–coupled receptor signaling. This study establishes a function for GIT2 in linking chemotaxis and superoxide production in neutrophils and shows that loss of GIT2 in vivo leads to an immunodeficient state.

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Figure 1: Immune effects of Git2 deficiency.
Figure 2: Expression of Git1 and Git2.
Figure 3: Git2 in neutrophil chemotaxis.
Figure 4: Impaired GPCR-coupled direction sensing in Git2−/− neutrophils.
Figure 5: Misoriented hyperproduction of superoxide in Git2−/− neutrophils.
Figure 6: Activity of small GTPases.

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  • 02 June 2006

    In the version of this article initially published online, the word "in" is missing on page 3, column 2, line 21; this should read "normal phosphorylation of PAK1 in Git2+/− neutrophils (Fig. 4d)." Also, in the supplementary information, the Greek mu symbol is missing from the units. The errors have been corrected for all versions of the article.

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Acknowledgements

We thank M. Hiraishi, Y. Shibata and M. Iwahara for help; H.A. Popiel (Osaka University, Suita, Japan) for critical reading of the manuscript; and R.Y. Tsien (University of California, La Jolla, California) for monomeric red fluorescent protein. Supported by Grants-In-Aid from the Ministry of Education, Science, Sports and Culture of Japan (16370090 to H.S.).

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Authors and Affiliations

  1. Department of Molecular Biology, Osaka Bioscience Institute, Suita, 565-0874, Japan

    Yuichi Mazaki, Shigeru Hashimoto, Masaki Morishige, Ari Hashimoto, Atsuko Yamada, Jin-Min Nam & Hisataka Sabe

  2. Department of Pathology, Hyogo College of Medicine, Nishinomiya, 663-8501, Japan

    Tohru Tsujimura

  3. Department of Neurosurgery, School of Medicine, Oita University, Oita, 879-5593, Japan

    Masaki Morishige

  4. Department of Molecular Behavioral Biology, Osaka Bioscience Institute, Suita, 565-0874, Japan

    Kosuke Aritake

  5. Laboratory for Animal Resources and Genetic Engineering, Center for Developmental Biology, RIKEN Kobe, Kobe, 650-0047, Japan

    Hiroshi Kiyonari & Kazuki Nakao

  6. Graduate School of Biostudies, Kyoto University, Kyoto, 606-8502, Japan

    Hisataka Sabe

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Supplementary information

Supplementary Fig. 1

Targeting of the Git2 gene by homologous recombination (PDF 287 kb)

Supplementary Fig. 2

Requirement of GAP activity for GIT2 function. (PDF 474 kb)

Supplementary Fig. 3

Subcellular localization of Rac1 and Rac2. (PDF 2124 kb)

Supplementary Table 1

Blood parameters of Git2+/− and Git2−/− mice. (PDF 17 kb)

Supplementary Table 2

DNA sequence of PCR primers. (PDF 25 kb)

Supplementary Methods (PDF 118 kb)

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Mazaki, Y., Hashimoto, S., Tsujimura, T. et al. Neutrophil direction sensing and superoxide production linked by the GTPase-activating protein GIT2. Nat Immunol 7, 724–731 (2006). https://doi.org/10.1038/ni1349

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