Distinct stages in stress granule assembly and disassembly
- University of Colorado Boulder, United States
- Howard Hughes Medical Institute, University of Colorado, United States
Figures
Figure 1
Time course of stress granule assembly.
(A) Two models for stress granule assembly. In Model 1, an increase in a pool of untranslated mRNAs with bound proteins containing IDRs, could lead to the formation of a LLPS based on weak, dynamic …
Figure 2
Stress granule cores are stable in lysates early in granule assembly.
(A) GFP-G3BP1 U-2 OS cells and cell lysates following treatment with NaAsO2 for 15 min. Graph shows percent GFP-G3BP1 foci for cells (in vivo) and cell lysates (in vitro) and normalized to maximal …
Figure 3
Early and late stress granules contain a non-uniform substructure.
GFP-G3BP1 stress granules were stained with the stress granule marker PABP1 and imaged by deconvolution microscopy (DV) following NaAsO2 stress at different time points. GFP-G3BP1 was assessed in …
Figure 4
Similar GFP-G3BP1 stress granule core size during early and prolonged stress.
Mean of 3 Nanosight experiments ± standard deviation for GFP-G3BP1 mammalian stress granule cores isolated from U-2 OS cells stressed with NaAs02 for 15, 30, 60, or 120 min. Relative number of …
Figure 5
Similar GFP-G3BP1 stress granule dynamics during early and prolonged stress.
Granules shown prior to photobleaching, at 0 s, and at 145 s after photobleaching. Cells were treated for either 30, 60, 120 min with NaAsO2. Graph shows recovery curves as an average of 6 granules …
Figure 6
Temperature inversely affects kinetics of stress granule formation.
(A) Time-lapse microscopy of U-2 OS cells expressing GFP-G3BP1. U-2 OS cells imaged at respective temperatures in the presence of NaAsO2 stress for 1 hr using 100X objective. (B) Graphical …
Figure 7
1,6-Hexanediol disrupts many cellular structures in yeast and alters cellular morphology in HeLa cells.
(A) The localization of various GFP-tagged proteins is shown either without treatment (No Drug); in the presence of 10% 1,6-Hexanediol and 10 μg/mL Digitonin for 2 min or 30 min; or just 10 μg/mL …
Figure 8
Effect of 1,6-Hexanediol on stress granules and P-bodies in yeast.
(A) Localization of Pab1-GFP and Edc3-mCherry prior to treatment with (Pre-treat) or after 2 min or 10 min of 10 μg/mL Digitonin ±10% 1,6-Hexanediol following 15 min of glucose starvation. All cells …
Figure 9
1,6-Hexanediol induces formation of stress granule-like assemblies in HeLa cells.
(A) Affects of 1,6-Hexanediol on NaAsO2-induced stress granules in HeLa cells. HeLa cells were stressed with NaAsO2 for 60 min. Addition of 3.5% 1,6-Hexanediol in the presence of NaAsO2 was added at …
Figure 10
Stress granules induced by 1,6-Hexanediol are cyclohexamide sensitive, stable in lysates, and dynamic.
(A) 1,6-Hexanediol induced stress granules are cyclohexamide sensitive in yeast and U-2 OS cells. Yeast were grown in the presence of 10% 1,6-Hexanediol and 10 μg/mL Digitonin for 10 min or just 10 …
Figure 11
Stress granules disassemble through multiple discrete steps.
(A) Time-lapse imaging of stress granule disassembly in normal media following 60 min of NaAsO2 stress using a 100X objective. Intensity map represents relative gray scale intensity of zoomed inset. …
Figure 12
Model for distinct stages of stress granule assembly and disassembly
Possible steps in granule assembly and disassembly are shown. Untranslating mRNPs nucleate an early stable mRNP (core) complex and grow to rapidly include an early core/ shell (biphasic) assembly …
Videos
Video 1
Stress granule assembly during NaAsO2 stress.
U-2 OS cells expressing GFP-G3BP1 were exposed to NaAsO2 (0.5 mM) and imaged for 20 min. Images were acquired at 20 s intervals on a spinning disk confocal microscope using a 100X objective. Scale …
Video 2
HeLa cells exposed to 1,6-Hexanediol.
HeLa cells were exposed to 1,6-Hexanediol (3.5%) and imaged for 5 min. Images were acquired at 10 s intervals on a spinning disk confocal using a 20X objective. Scale bar represents 20 µm. Related …
Video 3
Stress granule disassembly following NaAsO2 stress.
U-2 OS cells expressing GFP-G3BP1 stressed with NaAsO2 (0.5 mM) for 60 min and recovered in normal media. Image acquisition begins following 56 min of recovery in normal media. Images were acquired …
