Staurosporines Disrupt Phosphatidylserine Trafficking and Mislocalize Ras Proteins

Cho, Kwang Jin; Park, Jin Hee; Piggott, Andrew M.; Salim, Angela A.; Gorfe, Alemaheyu; Parton, Robert G.; Capon, Robert J.; Lacey, Ernest; Hancock, John F.

doi:10.1074/jbc.m112.424457

Cited by 91 publications

(151 citation statements)

References 50 publications

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“…An L(r)-r value above the 99% confidence interval demonstrates a nonrandom clustered distribution. Consistent with previous results, in wildtype (WT) cells we observed a nonrandom, clustered distribution of PS (Fairn et al, 2011;Cho et al, 2012; van der Hoeven Figure 2. Caveolin is essential for the regulation of K-Ras nanoclustering on the PM.…”

Section: Ras Nanoclusterssupporting

confidence: 80%

“…In consequence, H-and K-Ras assemble into spatially nonoverlapping nanoclusters with further lateral segregation into nonoverlapping GDP and GTP nanoclusters (Prior et al, 2003a;Plowman et al, 2005Plowman et al, , 2008Roy et al, 2005;Zhou et al, 2012). H-Ras associates with cholesterol-dependent nanoclusters on the inner leaflet of the PM when in an inactive GDP-bound state and, upon GTP loading, switches to cholesterol-insensitive nanodomains; K-Ras occupies cholesterol-insensitive nanoclusters in both GTP-and GDP-bound states that have been closely linked with phosphatidylserine (PS; Cho et al, 2012;van der Hoeven et al, 2013). Ras.GTP nanoclusters are the sole sites of effector recruitment and activation on the PM (Hibino et al, 2003;Murakoshi et al, 2004;Tian et al, 2007;Plowman et al, 2008), such that Ras signal transmission is abrogated if MEFs (n = 6).…”

Section: Cav1 Deficiency Is Associated With Changes In Membrane Lipidmentioning

confidence: 99%

“…Caveolae regulate the nanoscale organization of the plasma membrane to remotely control Ras signaling nanoclustering is inhibited, or nanocluster structure is perturbed Tian et al, 2007;Cho et al, 2012;Zhou et al, 2012;van der Hoeven et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

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Caveolae regulate the nanoscale organization of the plasma membrane to remotely control Ras signaling

Ariotti¹,

Zhou²,

Hill³

et al. 2014

J Gen Physiol

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Section: Ras Nanoclusterssupporting

confidence: 80%

Section: Cav1 Deficiency Is Associated With Changes In Membrane Lipidmentioning

confidence: 99%

See 1 more Smart Citation

Caveolae regulate the nanoscale organization of the plasma membrane to remotely control Ras signaling

Ariotti¹,

Zhou²,

Hill³

et al. 2014

J Gen Physiol

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“…Treating MDCK cells with staurosporine (STS) at low sub-apoptotic concentrations causes the relocalization of PtdSer from the plasma membrane to endosomes through an uncharacterized mechanism (Cho et al, 2012). Consistent with this finding, we found that treatment of CHO cells with STS led to the relocalization of the PtdSer biosensor GFP-Lact-C2 to endosomal compartments.…”

Section: Acute Alterations In Ptdser Distribution Alter Cholesterol Dsupporting

confidence: 77%

Complementary probes reveal that phosphatidylserine is required for the proper transbilayer distribution of cholesterol

Maekawa

Fairn

2015

Journal of Cell Science

234

249

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Cholesterol is an essential component of metazoan cellular membranes and it helps to maintain the structural integrity and fluidity of the plasma membrane. Here, we developed a cholesterol biosensor, termed D4H, based on the fourth domain of Clostridium perfringens theta-toxin, which recognizes cholesterol in the cytosolic leaflet of the plasma membrane and organelles. The D4H probe disassociates from the plasma membrane upon cholesterol extraction and after perturbations in cellular cholesterol trafficking. When used in combination with a recombinant version of the biosensor, we show that plasmalemmal phosphatidylserine is essential for retaining cholesterol in the cytosolic leaflet of the plasma membrane. In vitro experiments reveal that 1-stearoy-2-oleoyl phosphatidylserine can induce phase separation in cholesterol-containing lipid bilayers and shield cholesterol from cholesterol oxidase. Finally, the altered transbilayer distribution of cholesterol causes flotillin-1 to relocalize to endocytic organelles. This probe should be useful in the future to study pools of cholesterol in the cytosolic leaflet of the plasma membrane and organelles.

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“…Attempts to abrogate the plasma membrane binding of Ras, which is required for biological activity, by inhibiting farnesyl transferase (6,7) have failed because N-Ras and K-Ras are also good substrates for geranylgeranyl transferase 1 in cells treated with farnesyl transferase inhibitors (8,9). Other efforts along this line include the development of farnesyl analogs, currently in clinical trials (10), and other compounds that dislodge Ras from the plasma membrane (11,12). Although the potential therapeutic value and mechanism of action of these compounds are still under investigation, it is clear that they do not directly bind to Ras.…”

mentioning

confidence: 99%

Andrographolide derivatives inhibit guanine nucleotide exchange and abrogate oncogenic Ras function

Hocker

Cho

Chen

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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135

133

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Aberrant signaling by oncogenic mutant rat sarcoma (Ras) proteins occurs in ∼15% of all human tumors, yet direct inhibition of Ras by small molecules has remained elusive. Recently, several smallmolecule ligands have been discovered that directly bind Ras and inhibit its function by interfering with exchange factor binding. However, it is unclear whether, or how, these ligands could lead to drugs that act against constitutively active oncogenic mutant Ras. Using a dynamics-based pocket identification scheme, ensemble docking, and innovative cell-based assays, here we show that andrographolide (AGP)-a bicyclic diterpenoid lactone isolated from Andrographis paniculata-and its benzylidene derivatives bind to transient pockets on Kirsten-Ras (K-Ras) and inhibit GDP-GTP exchange. As expected for inhibitors of exchange factor binding, AGP derivatives reduced GTP loading of wild-type K-Ras in response to acute EGF stimulation with a concomitant reduction in MAPK activation. Remarkably, however, prolonged treatment with AGP derivatives also reduced GTP loading of, and signal transmission by, oncogenic mutant K-RasG12V. In sum, the combined analysis of our computational and cell biology results show that AGP derivatives directly bind Ras, block GDP-GTP exchange, and inhibit both wild-type and oncogenic K-Ras signaling. Importantly, our findings not only show that nucleotide exchange factors are required for oncogenic Ras signaling but also demonstrate that inhibiting nucleotide exchange is a valid approach to abrogating the function of oncogenic mutant Ras.cancer | molecular dynamics | allosteric site | drug design

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Staurosporines Disrupt Phosphatidylserine Trafficking and Mislocalize Ras Proteins

Cited by 91 publications

References 50 publications

Caveolae regulate the nanoscale organization of the plasma membrane to remotely control Ras signaling

Caveolae regulate the nanoscale organization of the plasma membrane to remotely control Ras signaling

Complementary probes reveal that phosphatidylserine is required for the proper transbilayer distribution of cholesterol

Andrographolide derivatives inhibit guanine nucleotide exchange and abrogate oncogenic Ras function

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