CoChat
Sign Up
Cited by 59Protein Kinase Regulation and GTPase SignalingCellular transport and secretionCellular Mechanics and Interactions

RHO binding to FAM65A regulates Golgi reorientation during cell migration

Open access
Mardakheh, Faraz K., Self, Annette, Marshall, Christopher J.·2016·Journal of Cell Science
DOI 10.1242/jcs.198614PMC PMC5201024
Open this paper in CoChat — free accountOpen PDF

Abstract

Directional cell migration involves reorientation of the secretory machinery. However, the molecular mechanisms that control this reorientation are not well characterised. Here, we identify a new Rho effector protein, named FAM65A, which binds to active RHOA, RHOB and RHOC. FAM65A links RHO proteins to Golgi-localising cerebral cavernous malformation-3 protein (CCM3; also known as PDCD10) and its interacting proteins mammalian STE20-like protein kinases 3 and 4 (MST3 and MST4; also known as STK24 and STK26, respectively). Binding of active RHO proteins to FAM65A does not affect the kinase activity of MSTs but results in their relocation from the Golgi in a CCM3-dependent manner. This relocation is crucial for reorientation of the Golgi towards the leading edge and subsequent directional cell migration. Our results reveal a previously unidentified pathway downstream of RHO that regulates the polarity of migrating cells through Golgi reorientation in a FAM65A-, CCM3- and MST3- and MST4-dependent manner.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1242/jcs.198614

Cites

2 of 2 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • The FAM65A/CCM3/MST relocation is essential for both membrane trafficking from the Golgi and orientation of the Golgi toward the leading edge of the cell.

    “This is essential for both membrane trafficking from the Golgi and the orientation of the Golgi toward the leading edge of the cell [128].”

    From RhoA Signaling in Neurodegenerative Diseases (2022) · cites this paper as [128]

    The Introduction and Discussion state that RHO-dependent relocation of the FAM65A–CCM3–MST complex is crucial for Golgi reorientation toward the leading edge. The Introduction discusses secretory traffic from the Golgi, but the paper does not establish that this complex’s relocation is essential for membrane trafficking from the Golgi. Page numbers are unavailable in our copy. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • Upon binding to RhoA, the FAM65A/CCM3/MST complex relocates away from the Golgi complex to the cytoplasmic membrane, relieving the inhibitory effect of MST4 on Golgi reorientation.

    “Upon binding to RhoA, the FAM65A/CCM3/MST complex relocates away from the Golgi complex to the cytoplasmic membrane, relieving the inhibitory effect of MST4 on Golgi reorientation.”

    From RhoA Signaling in Neurodegenerative Diseases (2022) · cites this paper as [128]

    The Discussion (page unavailable) proposes that RHO–GTP binding to FAM65A relocates the FAM65A–CCM3–MST complex from the Golgi, relieving MST-mediated inhibition of Golgi reorientation. The Results identify the destination as cytoplasmic punctae, likely vesicular compartments, not the ‘cytoplasmic membrane’ specified in the claim. (LLM, conf=0.87)

    Experimental — accuracy not yet measured

  • Activated RhoA binds FAM65A, which in turn binds CCM3 and MST.

    “Activated RhoA binds FAM65A, which in turn binds cerebral cavernous malformation-3 protein (CCM3) and the interacting mammalian STE20-like protein kinase (MST).”

    From RhoA Signaling in Neurodegenerative Diseases (2022) · cites this paper as [128]

  • FAM65A is a novel RhoA effector protein associated with the Golgi complex.

    “A novel RhoA effector protein FAM65A is also associated with the Golgi complex.”

    From RhoA Signaling in Neurodegenerative Diseases (2022) · cites this paper as [128]

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

Research works printed but not linked: 51

  1. Bauer D. E., Canver M. C. and Orkin S. H. (2015). Generation of genomic deletions in mammalian cell lines via CRISPR/Cas9. J. Vis. Exp. 95, e52118 10.3791/52118 PMC4279820 25549070 · DOI
  2. Bisel B., Wang Y., Wei J.-H., Xiang Y., Tang D., Miron-Mendoza M., Yoshimura S., Nakamura N. and Seemann J. (2008). ERK regulates Golgi and centrosome orientation towards the leading edge through GRASP65. J. Cell Biol. 182, 837- 843. 10.1083/jcb.200805045 18762583 PMC2528584 · DOI · Paper page
  3. Borikova A. L., Dibble C. F., Sciaky N., Welch C. M., Abell A. N., Bencharit S. and Johnson G. L. (2010). Rho kinase inhibition rescues the endothelial cell cerebral cavernous malformation phenotype. J. Biol. Chem. 285, 11760- 11764. 10.1074/jbc.C109.097220 20181950 PMC2852911 · DOI
  4. Bravi L., Rudini N., Cuttano R., Giampietro C., Maddaluno L., Ferrarini L., Adams R. H., Corada M., Boulday G., Tournier-Lasserve E. et al. (2015). Sulindac metabolites decrease cerebrovascular malformations in CCM3-knockout mice. Proc. Natl. Acad. Sci. USA 112, 8421- 8426. 10.1073/pnas.1501352112 26109568 PMC4500248 · DOI
  5. Ceccarelli D. F., Laister R. C., Mulligan V. K., Kean M. J., Goudreault M., Scott I. C., Derry W. B., Chakrabartty A., Gingras A. C. and Sicheri F. (2011). CCM3/PDCD10 heterodimerizes with germinal center kinase III (GCKIII) proteins using a mechanism analogous to CCM3 homodimerization. J. Biol. Chem. 286, 25056- 25064. 10.1074/jbc.M110.213777 21561863 PMC3137079 · DOI
  6. Coates J. C. (2003). Armadillo repeat proteins: beyond the animal kingdom. Trends Cell Biol. 13, 463- 471. 10.1016/S0962-8924(03)00167-3 12946625 · DOI
  7. Cox J. and Mann M. (2008). MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification. Nat. Biotechnol. 26, 1367- 1372. 10.1038/nbt.1511 19029910 · DOI · Paper page
  8. Cuttano R., Rudini N., Bravi L., Corada M., Giampietro C., Papa E., Morini M. F., Maddaluno L., Baeyens N., Adams R. H. et al. (2016). KLF4 is a key determinant in the development and progression of cerebral cavernous malformations. EMBO Mol. Med. 8, 6- 24. 10.15252/emmm.201505433 26612856 PMC4718159 · DOI
  9. Diaz-Horta O., Subasioglu-Uzak A., Grati M., Desmidt A., Foster J. II, Cao L., Bademci G., Tokgoz-Yilmaz S., Duman D., Cengiz F. B. et al. (2014). FAM65B is a membrane-associated protein of hair cell stereocilia required for hearing. Proc. Natl. Acad. Sci. USA 111, 9864- 9868. 10.1073/pnas.1401950111 24958875 PMC4103326 · DOI
  10. Draheim K. M., Fisher O. S., Boggon T. J. and Calderwood D. A. (2014). Cerebral cavernous malformation proteins at a glance. J. Cell Sci. 127, 701- 707. 10.1242/jcs.138388 24481819 PMC3924200 · DOI
  11. Etienne-Manneville S. (2004). Cdc42 - the centre of polarity. J. Cell Sci. 117, 1291- 1300. 10.1242/jcs.01115 15020669 · DOI
  12. Etienne-Manneville S. (2008). Polarity proteins in migration and invasion. Oncogene 27, 6970- 6980. 10.1038/onc.2008.347 19029938 · DOI
  13. Etienne-Manneville S. and Hall A. (2003). Cdc42 regulates GSK-3beta and adenomatous polyposis coli to control cell polarity. Nature 421, 753- 756. 10.1038/nature01423 12610628 · DOI · Paper page
  14. Etienne-Manneville S., Manneville J.-B., Nicholls S., Ferenczi M. A. and Hall A. (2005). Cdc42 and Par6-PKCzeta regulate the spatially localized association of Dlg1 and APC to control cell polarization. J. Cell Biol. 170, 895- 901. 10.1083/jcb.200412172 16157700 PMC2171429 · DOI
  15. Fidalgo M., Fraile M., Pires A., Force T., Pombo C. and Zalvide J. (2010). CCM3/PDCD10 stabilizes GCKIII proteins to promote Golgi assembly and cell orientation. J. Cell Sci. 123, 1274- 1284. 10.1242/jcs.061341 20332113 · DOI
  16. Flynn P., Mellor H., Palmer R., Panayotou G. and Parker P. J. (1998). Multiple interactions of PRK1 with RhoA. Functional assignment of the Hr1 repeat motif. J. Biol. Chem. 273, 2698- 2705. 10.1074/jbc.273.5.2698 9446575 · DOI
  17. Gao K., Tang W., Li Y., Zhang P., Wang D., Yu L., Wang C. and Wu D. (2015). Front-signal-dependent accumulation of the RHOA inhibitor FAM65B at leading edges polarizes neutrophils. J. Cell Sci. 128, 992- 1000. 10.1242/jcs.161497 25588844 PMC4342581 · DOI
  18. Gloerich M., Ten Klooster J. P., Vliem M. J., Koorman T., Zwartkruis F. J., Clevers H. and Bos J. L. (2012). Rap2A links intestinal cell polarity to brush border formation. Nat. Cell Biol. 14, 793- 801. 10.1038/ncb2537 22797597 · DOI
  19. Gordon J., Hwang J., Carrier K. J., Jones C. A., Kern Q. L., Moreno C. S., Karas R. H. and Pallas D. C. (2011). Protein phosphatase 2a (PP2A) binds within the oligomerization domain of striatin and regulates the phosphorylation and activation of the mammalian Ste20-Like kinase Mst3. BMC Biochem. 12, 54 10.1186/1471-2091-12-54 21985334 PMC3217859 · DOI
  20. Goudreault M., D'ambrosio L. M., Kean M. J., Mullin M. J., Larsen B. G., Sanchez A., Chaudhry S., Chen G. I., Sicheri F., Nesvizhskii A. I. et al. (2009). A PP2A phosphatase high density interaction network identifies a novel striatin-interacting phosphatase and kinase complex linked to the cerebral cavernous malformation 3 (CCM3) protein. Mol. Cell. Proteomics 8, 157- 171. 10.1074/mcp.M800266-MCP200 18782753 PMC2621004 · DOI · Paper page
  21. Infante C., Ramos-Morales F., Fedriani C., Bornens M. and Rios R. M. (1999). GMAP-210, A cis-Golgi network-associated protein, is a minus end microtubule-binding protein. J. Cell Biol. 145, 83- 98. 10.1083/jcb.145.1.83 10189370 PMC2148210 · DOI
  22. Kean M. J., Ceccarelli D. F., Goudreault M., Sanches M., Tate S., Larsen B., Gibson L. C. D., Derry W. B., Scott I. C., Pelletier L. et al. (2011). Structure-function analysis of core STRIPAK Proteins: a signaling complex implicated in Golgi polarization. J. Biol. Chem. 286, 25065- 25075. 10.1074/jbc.M110.214486 21561862 PMC3137080 · DOI
  23. Kümper S., Mardakheh F. K., Mccarthy A., Yeo M., Stamp G. W., Paul A., Worboys J., Sadok A., Jørgensen C., Guichard S. et al. (2016). Rho-associated kinase (ROCK) function is essential for cell cycle progression, senescence and tumorigenesis. eLife 5, e12203 10.7554/eLife.12203 26765561 PMC4798951 · DOI
  24. Maddaluno L., Rudini N., Cuttano R., Bravi L., Giampietro C., Corada M., Ferrarini L., Orsenigo F., Papa E., Boulday G. et al. (2013). EndMT contributes to the onset and progression of cerebral cavernous malformations. Nature 498, 492- 496. 10.1038/nature12207 23748444 · DOI · Paper page
  25. Madsen C. D., Hooper S., Tozluoglu M., Bruckbauer A., Fletcher G., Erler J. T., Bates P. A., Thompson B. and Sahai E. (2015). STRIPAK components determine mode of cancer cell migration and metastasis. Nat. Cell Biol. 17, 68- 80. 10.1038/ncb3083 25531779 PMC5354264 · DOI
  26. Marchler-Bauer A., Derbyshire M. K., Gonzales N. R., Lu S., Chitsaz F., Geer L. Y., Geer R. C., He J., Gwadz M., Hurwitz D. I. et al. (2015). CDD: NCBI's conserved domain database. Nucleic Acids Res. 43, D222- D226. 10.1093/nar/gku1221 25414356 PMC4383992 · DOI
  27. Mardakheh F. K., Auciello G., Dafforn T. R., Rappoport J. Z. and Heath J. K. (2010). Nbr1 is a novel inhibitor of ligand-mediated receptor tyrosine kinase degradation. Mol. Cell. Biol. 30, 5672- 5685. 10.1128/MCB.00878-10 20937771 PMC3004269 · DOI
  28. Mardakheh F. K., Paul A., Kümper S., Sadok A., Paterson H., Mccarthy A., Yuan Y. and Marshall C. J. (2015). Global analysis of mRNA, translation, and protein localization: local translation is a key regulator of cell protrusions. Dev. Cell 35, 344- 357. 10.1016/j.devcel.2015.10.005 26555054 PMC4643311 · DOI
  29. Matsui T., Yonemura S., Tsukita S. and Tsukita S. (1999). Activation of ERM proteins in vivo by Rho involves phosphatidyl-inositol 4-phosphate 5-kinase and not ROCK kinases. Curr. Biol. 9, 1259- 1262. 10.1016/S0960-9822(99)80508-9 10556088 · DOI
  30. Mellor H. (2004). Cell motility: Golgi signalling shapes up to ship out. Curr. Biol. 14, R434- R435. 10.1016/j.cub.2004.05.038 15182693 · DOI
  31. Palazzo A. F., Joseph H. L., Chen Y.-J., Dujardin D. L., Alberts A. S., Pfister K. K., Vallee R. B. and Gundersen G. G. (2001). Cdc42, dynein, and dynactin regulate MTOC reorientation independent of Rho-regulated microtubule stabilization. Curr. Biol. 11, 1536- 1541. 10.1016/S0960-9822(01)00475-4 11591323 · DOI
  32. Preisinger C., Short B., De Corte V., Bruyneel E., Haas A., Kopajtich R., Gettemans J. and Barr F. A. (2004). YSK1 is activated by the Golgi matrix protein GM130 and plays a role in cell migration through its substrate 14-3-3zeta. J. Cell Biol. 164, 1009- 1020. 10.1083/jcb.200310061 15037601 PMC2172068 · DOI
  33. Renz M., Otten C., Faurobert E., Rudolph F., Zhu Y., Boulday G., Duchene J., Mickoleit M., Dietrich A.-C., Ramspacher C. et al. (2015). Regulation of beta1 integrin-Klf2-mediated angiogenesis by CCM proteins. Dev. Cell 32, 181- 190. 10.1016/j.devcel.2014.12.016 25625207 · DOI
  34. Richardson B. T., Dibble C. F., Borikova A. L. and Johnson G. L. (2013). Cerebral cavernous malformation is a vascular disease associated with activated RhoA signaling. Biol. Chem. 394, 35- 42. 10.1515/hsz-2012-0243 23096573 PMC3677706 · DOI
  35. Ridley A. J. (2013). RhoA, RhoB and RhoC have different roles in cancer cell migration. J. Microsc. 251, 242- 249. 10.1111/jmi.12025 23488932 · DOI
  36. Ridley A. J., Self A. J., Kasmi F., Paterson H. F., Hall A., Marshall C. J. and Ellis C. (1993). rho family GTPase activating proteins p190, bcr and rhoGAP show distinct specificities in vitro and in vivo. EMBO J. 12, 5151- 5160. 8262058 PMC413777
  37. Rougerie P., Largeteau Q., Megrelis L., Carrette F., Lejeune T., Toffali L., Rossi B., Zeghouf M., Cherfils J., Constantin G. et al. (2013). Fam65b is a new transcriptional target of FOXO1 that regulates RhoA signaling for T lymphocyte migration. J. Immunol. 190, 748- 755. 10.4049/jimmunol.1201174 23241886 · DOI
  38. Sadok A. and Marshall C. J. (2014). Rho GTPases: masters of cell migration. Small GTPases 5, e983878 10.4161/sgtp.29710 PMC4107589 24978113 · DOI
  39. Sadok A., Mccarthy A., Caldwell J., Collins I., Garrett M. D., Yeo M., Hooper S., Sahai E., Kuemper S., Mardakheh F. K. et al. (2015). Rho kinase inhibitors block melanoma cell migration and inhibit metastasis. Cancer Res. 75, 2272- 2284. 10.1158/0008-5472.CAN-14-2156 25840982 · DOI
  40. Sahai E. and Olson M. F. (2006). Purification of TAT-C3 exoenzyme. Methods Enzymol. 406, 128- 140. 10.1016/S0076-6879(06)06011-3 16472655 · DOI
  41. Shaw R. J., Henry M., Solomon F. and Jacks T. (1998). RhoA-dependent phosphorylation and relocalization of ERM proteins into apical membrane/actin protrusions in fibroblasts. Mol. Biol. Cell 9, 403- 419. 10.1091/mbc.9.2.403 9450964 PMC25270 · DOI
  42. Stockton R. A., Shenkar R., Awad I. A. and Ginsberg M. H. (2010). Cerebral cavernous malformations proteins inhibit Rho kinase to stabilize vascular integrity. J. Exp. Med. 207, 881- 896. 10.1084/jem.20091258 20308363 PMC2856024 · DOI
  43. Ten Klooster J. P., Jansen M., Yuan J., Oorschot V., Begthel H., Di Giacomo V., Colland F., De Koning J., Maurice M. M., Hornbeck P. et al. (2009). Mst4 and Ezrin induce brush borders downstream of the Lkb1/Strad/Mo25 polarization complex. Dev. Cell 16, 551- 562. 10.1016/j.devcel.2009.01.016 19386264 · DOI
  44. Thumkeo D., Watanabe S. and Narumiya S. (2013). Physiological roles of Rho and Rho effectors in mammals. Eur. J. Cell Biol. 92, 303- 315. 10.1016/j.ejcb.2013.09.002 24183240 · DOI
  45. Tyanova S., Temu T., Sinitcyn P., Carlson A., Hein M. Y., Geiger T., Mann M. and Cox J. (2016). The Perseus computational platform for comprehensive analysis of (prote)omics data. Nat. Methods 13, 731- 740. 10.1038/nmeth.3901 27348712 · DOI
  46. Vizcaíno J. A., Deutsch E. W., Wang R., Csordas A., Reisinger F., Ríos D., Dianes J. A., Sun Z., Farrah T., Bandeira N. et al. (2014). ProteomeXchange provides globally coordinated proteomics data submission and dissemination. Nat. Biotechnol. 32, 223- 226. 10.1038/nbt.2839 24727771 PMC3986813 · DOI
  47. Whitehead K. J., Chan A. C., Navankasattusas S., Koh W., London N. R., Ling J., Mayo A. H., Drakos S. G., Jones C. A., Zhu W. et al. (2009). The cerebral cavernous malformation signaling pathway promotes vascular integrity via Rho GTPases. Nat. Med. 15, 177- 184. 10.1038/nm.1911 19151728 PMC2767168 · DOI
  48. Yadav S. and Linstedt A. D. (2011). Golgi positioning. Cold Spring Harb. Perspect. Biol. 3, a005322 10.1101/cshperspect.a005322 PMC3101843 21504874 · DOI
  49. Yadav S., Puri S. and Linstedt A. D. (2009). A primary role for Golgi positioning in directed secretion, cell polarity, and wound healing. Mol. Biol. Cell 20, 1728- 1736. 10.1091/mbc.E08-10-1077 19158377 PMC2655245 · DOI
  50. Zhou J., Shao Z., Kerkela R., Ichijo H., Muslin A. J., Pombo C. and Force T. (2009). Serine 58 of 14-3-3zeta is a molecular switch regulating ASK1 and oxidant stress-induced cell death. Mol. Cell. Biol. 29, 4167- 4176. 10.1128/MCB.01067-08 19451227 PMC2715813 · DOI
  51. Zhou Z., Tang A. T., Wong W.-Y., Bamezai S., Goddard L. M., Shenkar R., Zhou S., Yang J., Wright A. C., Foley M. et al. (2016). Cerebral cavernous malformations arise from endothelial gain of MEKK3–KLF2/4 signalling. Nature 532, 122- 126. 10.1038/nature17178 27027284 PMC4864035 · DOI

Cited by

Sorted by citations

Authors
Mardakheh, F. K., Self, A., & Marshall, C. J.
Affiliations
Institute of Cancer Research (GB)
Corresponding author
Self, Annette
ORCID
Venue
Journal of Cell Science
Publisher
The Company of Biologists
Published
2016-12-15
Volume
129
Issue
24
Pages
4466-4479
ISSN
  • 0021-9533 (ISSN-L)
  • 1477-9137
DOI
10.1242/jcs.198614
PMC
PMC5201024
OpenAlex
W2546935888
Cited by
59
References
2
Citations by year
2026: 6 · 2025: 6 · 2024: 2 · 2023: 6 · 2022: 8 · 2021: 13 · 2020: 8 · 2019: 3 · 2018: 4 · 2017: 2 · 2016: 1
FWCI
1.42
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Cell Science (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Queen Mary Research Online (Queen Mary University of London) (submittedVersion)
  • HighWire Press Open Archive (submittedVersion)
Funders
Cancer Research UK (C107/A12057, C107/A10433, C107/A16512)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
Keywords
Rho GTPases, RhoA, RhoB, directional cell migration, CCM3, PDCD10, MST3, MST4, secretory machinery, cell polarity, leading edge
MeSH terms
Hippo Kinases, Amino Acid Sequence, Cell Movement, Enzyme Activation, Golgi Apparatus, HeLa Cells, Humans, Membrane Proteins, Protein Binding, Proto-Oncogene Proteins, Protein Serine-Threonine Kinases, rho GTP-Binding Proteins, Protein Transport, Intracellular Signaling Peptides and Proteins, Apoptosis Regulatory Proteins

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review