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Cited by 13Mitochondrial Function and PathologyAutophagy in Disease and TherapyCellular transport and secretion

RalA and RalB relocalization to depolarized mitochondria depends on clathrin-mediated endocytosis and facilitates TBK1 activation

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Pollock, Sarah R., Schinlever, Austin R., Rohani, Ali, Kashatus, Jennifer A., Kashatus, David F.·2019·PLOS ONE
DOI 10.1371/journal.pone.0214764PMC PMC6469766
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Abstract

Healthy mitochondria use an electrochemical gradient across the inner mitochondrial membrane (IMM) to generate energy in the form of ATP. A variety of endogenous and exogenous factors can lead to transient or sustained depolarization of the IMM, including mitochondrial fission events, expression of uncoupling proteins, electron transport chain (ETC) inhibitors, or chemical uncouplers. This depolarization in turn leads to a variety of physiological responses, ranging from selective mitochondrial clearance (mitophagy) to cell death. How cells recognize and ultimately respond to depolarized mitochondria remains incompletely understood. Here we show that the small GTPases RalA and RalB both relocalize to mitochondria following depolarization in a process dependent on clathrin-mediated endocytosis (CME). Furthermore, both genetic and pharmacologic inhibition of RalA and RalB leads to an increase in the activity of the atypical IκB kinase TBK1 both basally and in response to mitochondrial depolarization. This phenotype was also observed following inhibition of Ral relocalization. Collectively, these data suggest a model in which RalA and RalB inhibit TBK1 and that relocalization of Ral to depolarized mitochondria facilitates TBK1 activation through release of this inhibition.

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  • RalA and RalB promote clearance of dysfunctional mitochondria by mitophagy; following depolarization they relocalize to mitochondria and activate TBK1.

    “It has also been found that RalA and RalB promote the clearance of dysfunctional mitochondria from the cell by mitophagy. Following the depolarization of a mitochondria, RalA and RalB relocalize to the mitochondria where they activate TBK1 which in turn promotes mitophagy [50].”

    From Rlip76: An Unexplored Player in Neurodegeneration and Alzheimer’s Disease? (2022) · cites this paper as [50]

    The Results section (‘RalA and RalB negatively regulate TBK1 activity’) reports that RalA and RalB relocalize after depolarization and that this relieves their inhibition of TBK1, allowing its activity to increase. The Introduction says Ral may be involved in mitophagy, but the paper does not show that RalA and RalB promote clearance of dysfunctional mitochondria by mitophagy. It also describes release of TBK1 inhibition, rather than direct activation by RalA and RalB. (LLM, conf=0.90)

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Research works printed but not linked: 42

  1. [1]Lim K-H, O'Hayer K, Adam SJ, Kendall SD, Campbell PM, Der CJ, et al Divergent roles for RalA and RalB in malignant growth of human pancreatic carcinoma cells. Current Biology. 2006 12 19; 16(24): 2385– 94. 10.1016/j.cub.2006.10.023 17174914 · DOI · Paper page
  2. [2]Smith SC, Oxford G, Baras AS, Owens C, Havaleshko D, Brautigan DL, et al Expression of ral GTPases, their effectors, and activators in human bladder cancer. Clin Cancer Res. Clinical Cancer Research; 2007 7 1; 13(13): 3803– 13. 10.1158/1078-0432.CCR-06-2419 17606711 · DOI
  3. [3]Martin TD, Samuel JC, Routh ED, Der CJ, Yeh JJ. Activation and involvement of Ral GTPases in colorectal cancer. Cancer Research. American Association for Cancer Research; 2011 1 1; 71(1): 206– 15. PMC3062918 21199803
  4. [4]Ginn KF. RalA is overactivated in medulloblastoma. J Neurooncol. Springer US; 2016 10 12; 130(1): 99– 110. 27566179
  5. [6]Peschard P, McCarthy A, Leblanc-Dominguez V, Yeo M, Guichard S, Stamp G, et al Genetic Deletion of RALA and RALB Small GTPases Reveals Redundant Functions in Development and Tumorigenesis. Current Biology. Elsevier Ltd; 2012 11 6; 22(21): 2063– 8. 23063435
  6. [7]Lim K-H, Baines AT, Fiordalisi JJ, Shipitsin M, Feig LA, Cox AD, et al Activation of RalA is critical for Ras-induced tumorigenesis of human cells. Cancer Cell. 2005 6; 7(6): 533– 45. 10.1016/j.ccr.2005.04.030 15950903 · DOI · Paper page
  7. [8]Kashatus DF, Lim K-H, Brady DC, Pershing NLK, Cox AD, Counter CM. RALA and RALBP1 regulate mitochondrial fission at mitosis. Nature Cell Biology. Nature Publishing Group; 2011 8 7; 13(8): 1– 10. PMC3167028 21822277
  8. [9]Moskalenko S, Tong C, Rosse C, Mirey G, Formstecher E, Daviet L, et al Ral GTPases regulate exocyst assembly through dual subunit interactions. Journal of Biological Chemistry. American Society for Biochemistry and Molecular Biology; 2003 12 19; 278(51): 51743– 8. 14525976
  9. [10]Moskalenko S, Henry DO, Rosse C, Mirey G, Camonis JH, White MA. The exocyst is a Ral effector complex. Nature Cell Biology. Nature Publishing Group; 2002 1; 4(1): 66– 72. 11740492
  10. [11]Bodemann BO, Orvedahl A, Cheng T, Ram RR, Ou Y-H, Formstecher E, et al RalB and the Exocyst Mediate the Cellular Starvation Response by Direct Activation of Autophagosome Assembly. Cell. Elsevier Inc; 2011 1 21; 144(2): 253– 67. PMC3038590 21241894
  11. [12]Martin TD, Chen X-W, Kaplan REW, Saltiel AR, Walker CL, Reiner DJ, et al Ral and Rheb GTPase activating proteins integrate mTOR and GTPase signaling in aging, autophagy, and tumor cell invasion. Molecular Cell. 2014 1 23; 53(2): 209– 20. 10.1016/j.molcel.2013.12.004 24389102 PMC3955741 · DOI
  12. [13]Chien Y, Kim S, Bumeister R, Loo Y-M, Kwon SW, Johnson CL, et al RalB GTPase-mediated activation of the IkappaB family kinase TBK1 couples innate immune signaling to tumor cell survival. Cell. 2006 10 6; 127(1): 157– 70. 10.1016/j.cell.2006.08.034 17018283 · DOI
  13. [14]Simicek M, Lievens S, Laga M, Guzenko D, Aushev VN, Kalev P, et al The deubiquitylase USP33 discriminates between RALB functions in autophagy and innate immune response. Nature Cell Biology. 2013 10; 15(10): 1220– 30. 10.1038/ncb2847 24056301 · DOI
  14. [15]Uhm M, Bazuine M, Zhao P, Chiang S-H, Xiong T, Karunanithi S, et al Phosphorylation of the exocyst protein Exo84 by TBK1 promotes insulin-stimulated GLUT4 trafficking. Sci Signal. American Association for the Advancement of Science; 2017 3 21; 10(471): eaah5085. 28325821
  15. [16]Moore AS, Holzbaur ELF. Dynamic recruitment and activation of ALS-associated TBK1 with its target optineurin are required for efficient mitophagy. Proc Natl Acad Sci USA. 2016 6 14; 113(24): E3349– 58. 10.1073/pnas.1523810113 27247382 PMC4914160 · DOI
  16. [17]Nuytemans K, Theuns J, Cruts M, Van Broeckhoven C. Genetic etiology of Parkinson disease associated with mutations in the SNCA, PARK2, PINK1, PARK7, and LRRK2 genes: a mutation update. Hum Mutat. Wiley Subscription Services, Inc., A Wiley Company; 2010 7; 31(7): 763– 80. PMC3056147 20506312
  17. [18]Dupuis L, Gonzalez de Aguilar J-L, Oudart H, de Tapia M, Barbeito L, Loeffler J-P. Mitochondria in amyotrophic lateral sclerosis: a trigger and a target. Neurodegener Dis. Karger Publishers; 2004; 1(6): 245– 54. 16908975
  18. [19]Twig G, Shirihai OS. The interplay between mitochondrial dynamics and mitophagy. Antioxid Redox Signal. 2011 5 15; 14(10): 1939– 51. 10.1089/ars.2010.3779 21128700 PMC3078508 · DOI · Paper page
  19. [20]Mouli PK, Twig G, Shirihai OS. Frequency and Selectivity of Mitochondrial Fusion Are Key to Its Quality Maintenance Function. Biophysj. Biophysical Society; 2009 5 6; 96(9): 3509– 18. PMC2711405 19413957
  20. [21]Dugast M, Toussaint H, Dousset C, Benaroch P. AP2 clathrin adaptor complex, but not AP1, controls the access of the major histocompatibility complex (MHC) class II to endosomes. Journal of Biological Chemistry. 2005 5 20; 280(20): 19656– 64. 10.1074/jbc.M501357200 15749704 · DOI
  21. [22]Lim K-H, Brady DC, Kashatus DF, Ancrile BB, Der CJ, Cox AD, et al Aurora-A phosphorylates, activates, and relocalizes the small GTPase RalA. Molecular and Cellular Biology. American Society for Microbiology; 2010 1; 30(2): 508– 23. PMC2798468 19901077
  22. [23]Narendra D, Tanaka A, Suen DF, Youle RJ. Parkin is recruited selectively to impaired mitochondria and promotes their autophagy. The Journal of Cell Biology. 2008 11 24; 183(5): 795– 803. 10.1083/jcb.200809125 19029340 PMC2592826 · DOI · Paper page
  23. [24]Rainbolt TK, Lebeau J, Puchades C, Wiseman RL. Reciprocal Degradation of YME1L and OMA1 Adapts Mitochondrial Proteolytic Activity during Stress. CellReports. 2016 3 8; 14(9): 2041– 9. PMC4785047 26923599
  24. [25]Frech M, Schlichting I, Wittinghofer A, Chardin P. Guanine nucleotide binding properties of the mammalian RalA protein produced in Escherichia coli. Journal of Biological Chemistry. American Society for Biochemistry and Molecular Biology; 1990 4 15; 265(11): 6353– 9. 2108160
  25. [26]Emkey R, Freedman S, Feig LA. Characterization of a GTPase-activating protein for the Ras-related Ral protein. Journal of Biological Chemistry. 1991 5 25; 266(15): 9703– 6. 1903395
  26. [27]Jullien-Flores V, Mahé Y, Mirey G, Leprince C, Meunier-Bisceuil B, Sorkin A, et al RLIP76, an effector of the GTPase Ral, interacts with the AP2 complex: involvement of the Ral pathway in receptor endocytosis. Journal of Cell Science. 2000 8; 113 (Pt 16): 2837– 44. 10910768
  27. [28]Lim KH, Brady DC, Kashatus DF, Ancrile BB, Der CJ, Cox AD, et al Aurora-A phosphorylates, activates, and relocalizes the small GTPase RalA. Molecular and Cellular Biology. American Society for Microbiology; 2010 1 30; 30(2): 508– 23. PMC2798468 19901077
  28. [29]Bivona TG, Quatela SE, Bodemann BO, Ahearn IM, Soskis MJ, Mor A, et al PKC regulates a farnesyl-electrostatic switch on K-Ras that promotes its association with Bcl-XL on mitochondria and induces apoptosis. Molecular Cell. 2006 2 17; 21(4): 481– 93. 10.1016/j.molcel.2006.01.012 16483930 · DOI
  29. [30]Wu J-C, Chen T-Y, Yu C-TR, Tsai S-J, Hsu J-M, Tang M-J, et al Identification of V23RalA-Ser194 as a critical mediator for Aurora-A-induced cellular motility and transformation by small pool expression screening. Journal of Biological Chemistry. American Society for Biochemistry and Molecular Biology; 2005 3 11; 280(10): 9013– 22. 15637052
  30. [31]Wang H, Owens C, Chandra N, Conaway MR, Brautigan DL, Theodorescu D. Phosphorylation of RalB is important for bladder cancer cell growth and metastasis. Cancer Research. American Association for Cancer Research; 2010 11 1; 70(21): 8760– 9. PMC3025490 20940393
  31. [32]Sablina AA, Chen W, Arroyo JD, Corral L, Hector M, Bulmer SE, et al The tumor suppressor PP2A Abeta regulates the RalA GTPase. Cell. 2007 6 1; 129(5): 969– 82. 10.1016/j.cell.2007.03.047 17540176 PMC1945132 · DOI
  32. [33]Pollock SR, Kashatus DF. Ral In: Encyclopedia of Signaling Molecules. Cham: Springer, Cham; 2018 pp. 4424– 33. (Encyclopedia of Signaling Molecules; vol. 39).
  33. [34]Elkin SR, Lakoduk AM, Schmid SL. Endocytic pathways and endosomal trafficking: a primer Wien Med Wochenschr. Springer Vienna; 2016 2 9;: 1– 9 PMC4873410 26861668
  34. [35]Nakashima S, Morinaka K, Koyama S, Ikeda M, Kishida M, Okawa K, et al Small G protein Ral and its downstream molecules regulate endocytosis of EGF and insulin receptors. The EMBO Journal. EMBO Press; 1999 7 1; 18(13): 3629– 42. PMC1171441 10393179
  35. [37]Shen Y, Xu L, Foster DA. Role for phospholipase D in receptor-mediated endocytosis. Molecular and Cellular Biology. American Society for Microbiology; 2001 1; 21(2): 595– 602. PMC86627 11134345
  36. [38]Landry M-C, Champagne C, Boulanger M-C, Jetté A, Fuchs M, Dziengelewski C, et al A functional interplay between the small GTPase Rab11a and mitochondria-shaping proteins regulates mitochondrial positioning and polarization of the actin cytoskeleton downstream of Src family kinases. J Biol Chem. American Society for Biochemistry and Molecular Biology; 2014 1 24; 289(4): 2230– 49. PMC3900968 24302731
  37. [39]Hamacher-Brady A, Choe SC, Krijnse-Locker J, Brady NR. Intramitochondrial recruitment of endolysosomes mediates Smac degradation and constitutes a novel intrinsic apoptosis antagonizing function of XIAP E3 ligase. Cell Death and Differentiation. Nature Publishing Group; 2014 8 1; 21(12): 1862– 76. PMC4227142 25080938
  38. [41]Heo J-M, Ordureau A, Paulo JA, Rinehart J, Harper JW. The PINK1-PARKIN Mitochondrial Ubiquitylation Pathway Drives a Program of OPTN/NDP52 Recruitment and TBK1 Activation to Promote Mitophagy. Molecular Cell. Elsevier Inc; 2015 9 8;: 1– 15. PMC4592482 26365381
  39. [42]Neyraud V, Aushev VN, Hatzoglou A, Meunier B, Cascone I, Camonis J. RalA and RalB proteins are ubiquitinated GTPases, and ubiquitinated RalA increases lipid raft exposure at the plasma membrane. Journal of Biological Chemistry. American Society for Biochemistry and Molecular Biology; 2012 8 24; 287(35): 29397– 405. PMC3436142 22700969
  40. [43]Pilli M, Arko-Mensah J, Ponpuak M, Roberts E, Master S, Mandell MA, et al TBK-1 promotes autophagy-mediated antimicrobial defense by controlling autophagosome maturation. Immunity. 2012 8 24; 37(2): 223– 34. 22921120 PMC3428731
  41. [44]Collins LV, Hajizadeh S, Holme E, Jonsson I-M, Tarkowski A. Endogenously oxidized mitochondrial DNA induces in vivo and in vitro inflammatory responses. J Leukoc Biol. 2004 6; 75(6): 995– 1000. 10.1189/jlb.0703328 14982943 · DOI
  42. [45]West AP, Khoury-Hanold W, Staron M, Tal MC, Pineda CM, Lang SM, et al Mitochondrial DNA stress primes the antiviral innate immune response. Nature. 2015 2 2; 520(7548): 553– 7. 10.1038/nature14156 25642965 PMC4409480 · DOI · Paper page

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Authors
Pollock, S. R., Schinlever, A. R., Rohani, A., Kashatus, J. A., & Kashatus, D. F.
Affiliations
University of Virginia (US)
Corresponding author
  • Pollock, Sarah R.
  • Schinlever, Austin R.
  • Rohani, Ali
  • Kashatus, Jennifer A.
  • Kashatus, David F.
ORCID
Venue
PLOS ONE
Publisher
Public Library of Science (PLoS)
Published
2019-04-17
Volume
14
Issue
4
Pages
e0214764
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0214764
PMC
PMC6469766
OpenAlex
W2939228386
Cited by
13
References
3
Citations by year
2026: 2 · 2024: 1 · 2023: 1 · 2022: 2 · 2021: 2 · 2020: 4 · 2019: 1
FWCI
0.74
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Figshare (submittedVersion, cc-by)
Funders
National Cancer Institute (1R01CA200755-01)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
  • Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
Keywords
RalA, RalB, Ral GTPase, mitochondrial depolarization, clathrin-mediated endocytosis, IκB kinase, inner mitochondrial membrane, mitochondrial fission, mitophagy
MeSH terms
Carbonyl Cyanide m-Chlorophenyl Hydrazone, Clathrin, Endocytosis, HeLa Cells, Humans, Mitochondria, Models, Biological, Recombinant Proteins, Protein Serine-Threonine Kinases, ral GTP-Binding Proteins, Protein Transport, RNA, Small Interfering, Membrane Potential, Mitochondrial

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