ZFIN Image: Liao et al., 2019, Figure 1

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Figure Caption

Figure 1

RNA Granules Hitchhike on Motile Lysosomes in Mammalian Cells

(A) RNA granule co-imaging with different organelles. U2OS cells expressing mCherry-G3BP1 and different organelle markers were imaged live 30 minutes after heat shock (43^oC). Organelle markers: LAMP1 – lysosome, Sec61 – ER, TOMM20 – mitochondria, SiT – Golgi, SKL – peroxisome, Rab5 – early endosome, Rab7 – late endosome, Rab11a – recycling endosome, Ensconsin – microtubule. Arrows point to lysosome-RNA granule contact sites. Scale bar: 2μm. See also Figures S1A, S1B.

(B) Percentage of RNA granules that co-traffic with different organelles from (A). N=7.

(C) Time-lapse image sequence showing RNA granule (mCherry-G3BP1) co-trafficking with a lysosome (LAMP1-HaloTag) along a microtubule (Ensconsin-GFP) in U20S cells immediately after heat shock at 43^oC. Scale bar: 1μm. See also Figure S1F, Videos S1, S2.

(D) Kymograph of RNA granules co-trafficking with lysosomes in axons.

Axons of rat cortical neurons expressing LAMP1-HaloTag and mEmerald-G3BP1 were imaged at 100ms/frame for 30 seconds. Arrow points to a lysosome co-trafficking with a G3BP1-labeled structure. P50/p150Glued: Doxycycline-inducible expression of a p50 dynactin subunit and the CC1 domain of the p150 glued subunit of dynactin was used to inhibit motor-directed transport of lysosomes. Scale bar: 5 μm. See also Figures S1J, S1K.

(E) Kymograph of mRNA co-trafficking with lysosomes in axons.

Axons of rat cortical neurons expressing LAMP1-HaloTag, actin-24xMBS and MCP-NLS-2xEGFP were imaged as in (D). Arrow points to a lysosome co-trafficking with actin mRNA. Scale bar: 5 μm. See also Figure S1I.

(F) CLEM images of an RNA granule associated with a lysosome. Upper panel shows the fluorescent image of a LAMP1-labeled lysosome and a G3BP1-labeled RNA granule. Lower panel shows the correlated electron microscopy image. L, lysosome; G, RNA granule. Scale bar: 1μm.

Acknowledgments

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Reprinted from Cell, 179, Liao, Y.C., Fernandopulle, M.S., Wang, G., Choi, H., Hao, L., Drerup, C.M., Patel, R., Qamar, S., Nixon-Abell, J., Shen, Y., Meadows, W., Vendruscolo, M., Knowles, T.P.J., Nelson, M., Czekalska, M.A., Musteikyte, G., Gachechiladze, M.A., Stephens, C.A., Pasolli, H.A., Forrest, L.R., St George-Hyslop, P., Lippincott-Schwartz, J., Ward, M.E., RNA Granules Hitchhike on Lysosomes for Long-Distance Transport, Using Annexin A11 as a Molecular Tether, 147-164.e20, Copyright (2019) with permission from Elsevier. Full text @ Cell