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Published online
doi:10.1083/jcb.200604114
The Journal of Cell Biology, Vol. 178, No. 2, 245-255
The Rockefeller University Press, 0021-9525 $30.00
© Cho et al.
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Article

Localization of Gi{alpha} proteins in the centrosomes and at the midbody: implication for their role in cell division



Hyeseon Cho and John H. Kehrl

B-Cell Molecular Immunology Section, Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892

Correspondence to Hyeseon Cho: hcho{at}niaid.nih.gov; or John H. Kehrl: kehrl{at}niaid.nih.gov

At the plasma membrane, heterotrimeric G proteins act as molecular switches to relay signals from G protein–coupled receptors; however, G{alpha} subunits also have receptor-independent functions at intracellular sites. Regulator of G protein signaling (RGS) 14, which enhances the intrinsic GTPase activity of Gi{alpha} proteins, localizes in centrosomes, which suggests the coexpression of Gi{alpha}. We show expression of Gi{alpha}1, Gi{alpha}2, and Gi{alpha}3 in the centrosomes and at the midbody. Fluorescence resonance energy transfer analysis confirms a direct interaction between RGS14 and Gi{alpha}1 in centrosomes. Expression of GTPase-deficient Gi{alpha}1 results in defective cytokinesis, whereas that of wild-type or GTPase-deficient Gi{alpha}3 causes prolonged mitosis. Cells treated with pertussis toxin, with reduced expression of Gi{alpha}1, Gi{alpha}2, and Gi{alpha}3 or with decreased expression of RGS14 also exhibit cytokinesis defects. These results suggest that Gi{alpha} proteins and their regulators at these sites may play essential roles during mammalian cell division.

Abbreviations list: DIC, differential interference contrast; EE, Glu-Glu; FRET, fluorescence resonance energy transfer; GAP, GTPase-activating protein; GDI, guanine nucleotide dissociation inhibitor; GEF, guanine nucleotide exchange factor; GPCR, G protein–coupled receptor; MT, microtubule; PTX, pertussis toxin; RGS, regulator of G protein signaling.


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