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Calcium-dependent interaction of Lis1 with IQGAP1 and Cdc42 promotes neuronal motility

Abstract

Lis1 gene defects impair neuronal migration, causing the severe human brain malformation lissencephaly. Although much is known about its interactions with microtubules, microtubule-binding proteins such as CLIP-170, and with the dynein motor complex, the response of Lis1 to neuronal motility signals has not been elucidated. Lis1 deficiency is associated with deregulation of the Rho-family GTPases Cdc42, Rac1 and RhoA, and ensuing actin cytoskeletal defects, but the link between Lis1 and Rho GTPases remains unclear. We report here that calcium influx enhances neuronal motility through Lis1-dependent regulation of Rho GTPases. Lis1 promotes Cdc42 activation through interaction with the calcium sensitive GTPase scaffolding protein IQGAP1, maintaining the perimembrane localization of IQGAP1 and CLIP170 and thereby tethering microtubule ends to the cortical actin cytoskeleton. Lis1 thus is a key component of neuronal motility signal transduction that regulates the cytoskeleton by complexing with IQGAP1, active Cdc42 and CLIP-170 upon calcium influx.

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Figure 1: D-Serine requires Lis1 to increase neuronal migration in a Cdc42-dependent manner.
Figure 2: D-Serine activation of Rho GTPases is impaired in Lis1+/− neurons.
Figure 3: D-Serine and Lis1 regulate F-actin content in a Cdc42-dependent manner.
Figure 4: Lis1 levels modulate IQGAP1 and CLIP-170 distribution.
Figure 5: Ca2+-dependent redistribution of CLIP-170 and IQGAP1 to the insoluble fraction is impaired in Lis1+/− neurons.
Figure 6: Lis1 and IQGAP1 colocalize in cells and coprecipitate with WASP-CD agarose (WCD) along with Cdc42-GTP, and by direct immunoprecipitation.
Figure 7: RNAi-targeted decreases in CLIP-170 or IQGAP1 reduce Lis1 and Cdc42 in WCD-complexes.

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Acknowledgements

Supported by the US National Institute of Neurological Disorders and Stroke (RO1NS35515 to M.E.R., PO1NS39404 to M.E.R. and A.W.B.) and National Institute of Child Health and Development (RO1HD19950 to P.C.L.).

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Correspondence to M Elizabeth Ross.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

D-Serine fails to stimulate Lis1+/− neuronal migration despite enhancing Ca2+ fluctuations. (PDF 158 kb)

Supplementary Fig. 2

Bioactivity of plasmids expressing constitutively active G12V-Cdc42 (Cdc42ca) or dominant negative T17N-Cdc42 (Cdc42dn). (PDF 722 kb)

Supplementary Fig. 3

Demonstration of IQGAP1 down-regulation by shRNAi. (PDF 164 kb)

Supplementary Fig. 4

Demonstration of CLIP-170 down-regulation by shRNAi. (PDF 178 kb)

Supplementary Fig. 5

Model for Lis1 role in Ca2+ modulation of Rho GTPases and neuronal cytoskeleton. (PDF 816 kb)

Supplementary Fig. 6

Phosphorylation of Lis1 is enhanced by D-serine treatment. (PDF 147 kb)

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Kholmanskikh, S., Koeller, H., Wynshaw-Boris, A. et al. Calcium-dependent interaction of Lis1 with IQGAP1 and Cdc42 promotes neuronal motility. Nat Neurosci 9, 50–57 (2006). https://doi.org/10.1038/nn1619

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