Differential Localization of Rho Gtpases in Live Cells
By D. Michaelson, Joseph Silletti, G. Murphy, P. D’Eustachio, M. Rush, M. Philips. Journal of Cell Biology, 2001
745 citations recorded by the source corpus.
Determinants of membrane targeting of Rho proteins were investigated in live cells with green fluorescent fusion proteins expressed with or without Rho-guanine nucleotide dissociation inhibitor (GDI)α. The hypervariable region determined to which membrane compartment each protein was targeted. Targeting was regulated by binding to RhoGDIα in the case of RhoA, Rac1, Rac2, and Cdc42hs but not RhoB or TC10. Although RhoB localized to the plasma membrane (PM), Golgi, and motile peri-Golgi vesicles, TC10 localized to PMs and endosomes. Inhibition of palmitoylation mislocalized H-Ras, RhoB, and TC10 to the endoplasmic reticulum. Although overexpressed Cdc42hs and Rac2 were observed predominantly on endomembrane, Rac1 was predominantly at the PM. RhoA was cytosolic even when expressed at levels in vast excess of RhoGDIα. Oncogenic Dbl stimulated translocation of green fluorescent protein (GFP)-Rac1, GFP-Cdc42hs, and GFP-RhoA to lamellipodia. RhoGDI binding to GFP-Cdc42hs was not affected by substituting farnesylation for geranylgeranylation. A palmitoylation site inserted into RhoA blocked RhoGDIα binding. Mutations that render RhoA, Cdc42hs, or Rac1, either constitutively active or dominant negative abrogated binding to RhoGDIα and redirected expression to both PMs and internal membranes. Thus, despite the common essential feature of the CAAX (prenylation, AAX tripeptide proteolysis, and carboxyl methylation) motif, the subcellular localizations of Rho GTPases, like their functions, are diverse and dynamic.
Subfields: Cell Biology, Molecular Biology
View Paper: https://www.semanticscholar.org/paper/2d1e0349b07eef45f679edd4df66b2e1d5546ef0
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In Cell Biology
- A. Ridley
- A. Hall
- C. Der
- Sandrine Étienne-Manneville
- Krister Wennerberg
- S. Heasman
- K. Rossman
- G. Gundersen
In Molecular Biology
- A. Ridley
- C. Der
- Krister Wennerberg
- A. Hall
- Sandrine Étienne-Manneville
- S. Heasman
- K. Rossman
- P. Chiarugi
Citations in the corpus
- The complex of Arl2‐GTP and PDEδ: from structure to function (2002, result): “Instead, our result points to a preference of PDEd for H-Ras-GDP over H-Ras-GTP, which is in agreement with observations made with RhoGDI (Michaelson et al., 2001).”
- Identification and Characterization of MAVS, a Mitochondrial Antiviral Signaling Protein that Activates NF-κB and IRF3 (2005, neutral): “These sequences include the CAAX motif from Rac1 that is known to target fusion proteins predominantly to the plasma membrane (Michaelson et al., 2001); the ER targeting sequence from VAMP-2 (vesicle associated membrane protein-2), which is predominantly an ER membrane protein (Kim et al.”
- Integrin-mediated activation of Cdc42 controls cell polarity in migrating astrocytes through PKCzeta. (2001, background): “To identify the atypical PKC involved, we expressed a kinase-dead version of PKC in cells, and this blocked that GFP-Cdc42 localizes to the same cellular location as endogenous Cdc42 (Michaelson et al., 2001).”
- Rho-family GTPases: it's not only Rac and Rho (and I like it) (2004, background): “RhoA and RhoB localize very differently in the cell (Gampel et al., 1999; Michaelson et al., 2001), potentially because of their differential GDI binding.”
- Rho GTPases and cell migration. (2001, background): “Interestingly, Cdc42 localizes predominantly to the Golgi complex, although it is also present on the plasma membrane (Erickson et al., 1996), and possibly on other intracellular vesicles (Michaelson et al., 2001).”