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Optogenetic oligomerization of Rab GTPases regulates intracellular membrane trafficking

Nature Chemical Biology volume 12, pages 431–436 (2016)Cite this article

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Abstract

Intracellular membrane trafficking, which is involved in diverse cellular processes, is dynamic and difficult to study in a spatiotemporal manner. Here we report an optogenetic strategy, termed light-activated reversible inhibition by assembled trap of intracellular membranes (IM-LARIAT), that uses various Rab GTPases combined with blue-light-induced hetero-interaction between cryptochrome 2 and CIB1. In this system, illumination induces a rapid and reversible intracellular membrane aggregation that disrupts the dynamics and functions of the targeted membrane. We applied IM-LARIAT to specifically perturb several Rab-mediated trafficking processes, including receptor transport, protein sorting and secretion, and signaling initiated from endosomes. We finally used this tool to reveal different functions of local Rab5-mediated and Rab11-mediated membrane trafficking in growth cones and soma of young hippocampal neurons. Our results show that IM-LARIAT is a versatile tool that can be used to dissect spatiotemporal functions of intracellular membranes in diverse systems.

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Figure 1: Optogenetic control of intracellular membrane trafficking by light-induced aggregation of Rab GTPases.
Figure 2: Optogenetic control of specific functions mediated by Rab5-targeted and Rab11-targeted membrane.
Figure 3: Optogenetic control of specific functions mediated by diverse Rab-targeted membranes.
Figure 4: Rab5-targeted and Rab11-targeted aggregations in GCs of hippocampal neurons differently interfere with protrusion and growth.

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Acknowledgements

We thank N. Kim, H. Lee and M. Lee for helpful discussions, and D. Woo, H. Jung and S. Kim for technical assistance in neuron culture. This work was supported by the Institute for Basic Science (no. IBS-R001-G1); KAIST Institute for the BioCentury, Republic of Korea; and the Intelligent Synthetic Biology Center of the Global Frontier Project (2011-0031955) funded by the Ministry of Science, ICT and Future Planning, Republic of Korea.

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Authors and Affiliations

  1. Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Republic of Korea

    Mai Khanh Nguyen, Hyerim Park & Won Do Heo

  2. Graduate School of Nanoscience and Technology, KAIST, Daejeon, Republic of Korea

    Cha Yeon Kim

  3. Graduate School of Medical Science and Engineering, KAIST, Daejeon, Republic of Korea

    Jin Man Kim

  4. Center for Cognition and Sociality, Institute for Basic Science (IBS), Daejeon, Republic of Korea

    Byung Ouk Park, Sangkyu Lee & Won Do Heo

  5. KAIST Institute for the BioCentury, KAIST, Daejeon, Republic of Korea

    Won Do Heo

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  1. Mai Khanh Nguyen
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Contributions

M.K.N. and W.D.H. conceived the idea. M.K.N. and C.Y.K. designed and performed the experiments. M.K.N., C.Y.K. and J.M.K. analyzed the data. M.K.N., C.Y.K., J.M.K., B.O.P., S.L., H.P. and W.D.H. discussed the data. M.K.N. and W.D.H. wrote the paper.

Corresponding author

Correspondence to Won Do Heo.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Results and Supplementary Figures 1–11. (PDF 3556 kb)

Expression of CRY2-conjugated Rab GTPase results in an aberrant vesicle pattern and slow dynamics. (MOV 4186 kb)

Optogenetic control of early and recycling endosomes. (MOV 4970 kb)

Optogenetic control of late endosome trafficking. (MOV 1563 kb)

Optogenetic control of secretory vesicle trafficking. (MOV 4622 kb)

Optogenetic control of ER-Golgi-PM vesicle trafficking. (MOV 3718 kb)

Rab5-targeted aggregation in NIH-3T3 cells abrogates PDGF-induced circular dorsal ruffle formation. (MOV 1685 kb)

Local Rab5-targeted aggregation in rat hippocampal neurons. (MOV 8974 kb)

Local Rab5- and Rab11-targeted aggregation in the soma does not affect protrusion rate of growth cones. (MOV 3679 kb)

Local Rab5- and Rab11-targeted aggregation in the growth cones induce reduction in growth cone area. (MOV 1138 kb)

Local Rab5-targeted aggregation in the fast-protruding growth cones interferes with protrusion rate. (MOV 1697 kb)

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Nguyen, M., Kim, C., Kim, J. et al. Optogenetic oligomerization of Rab GTPases regulates intracellular membrane trafficking. Nat Chem Biol 12, 431–436 (2016). https://doi.org/10.1038/nchembio.2064

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