Publications

Starling GP, Philips BA, Ganesh S and King JS†. Katnip is required to maintain microtubule function and lysosomal delivery to autophagosomes and phagosomes. Molecular Biology of the Cell. 2023. 34 (3), ar12

Lutton JE, Coker H, Paschke P, Munn CJ, King JS†, Bretschneider T† and Kay RR†. Formation and closure of macropinocytic cups in Dictyostelium. Current Biology. 2023. 33 (15), 3083-3096. e6

Using 3D timelapse microscopy we provide new insights into how macropinosome formation. We find that cups close in two different ways and propose a new model for closure based on membrane tension. Commentary by Joel Swanson here.

Vines JH, Maib H, Buckley CM, Gueho A, Zhu Z, Soldati  T, Murray DH and King JS. A PI(3,5)P2 reporter reveals PIKfyve activity and dynamics on macropinosomes and phagosomes. The Journal of Cell Biology. 2023. 222 (9): e202209077.

We describe the first reliable reporter for PI(3,5)P2, allowing this lipid to be visualised in live cells for the first time. Commentary by Gerry Hammond here.

Kay RR, Lutton J, Coker H, Paschke P, JS King, Bretschneider T. The amoeba model for macropinocytosis. 2022. In: Commisso, C. (ed) Macropinocytosis. Subcellular Biochemistry, vol 98. Springer, Cham 

Itabangi H, Sephton-Clark PCS, Tamayo DP, Zhou X, Starling GP, Mahamoud Z, Insua I, Probert M, Correia J, Moynihan PJ, Gebremariam T, Yiyou G, Ibrahim AS, Brown GD, King JS†, Ballou ER†, Voelz K†. A bacterial endosymbiont of the fungus Rhizopus microsporus drives phagocyte evasion and opportunistic virulence. Current Biology, 2022. 32;5 1115-1130. 

Here, we show that a clinical fungal isolate harbours endocytic bacteria that are required for it's virulence. We show this is due to a secreted activity that inhibits phagocyte activities. As both fungus and bacteria normally live in the soil and only opportunistically infect humans we propose that endosymbiosis and virulence traits evolved to evade environmental predators, such as amoebae.

Vines JH, Buckley CM, Maib H, Gueho A, Soldati T, Murray D and King JS†. A PI(3,5)P2 probe reveals PIKfyve is required for Rab7 acquisition and the delivery and fusion of early macropinosomes to phagosomes. BioRxiv 2022.09. 19.508510

Williams SB, Chubb JR, Insall RHI, King JS, Pears CJ, Thompson E and Weijer CJ. Moving the research forward: the best of British biology using the tractable model system Dictyostelium Discoideum. Cells, 2021, 10(11), 3036

JS King. Dynamic Rac1 inhibition by CYRI helps cells drink, but stops them from driving. Journal of Cell Biology. 2021 220(9) DOI:  10.1083/jcb.202108041.  Commentary on new work by the Machesky lab

King JS and Smythe E. Water loss regulates cell and vesicle volume. Science, 2020. 367: 246-247

Buckley CM, Potts R, Gueho A, Vines JH, Munn CJ, Phillips BA, Gilsbach B, Nikolaev A, Soldati T, Parnell AJ, Kortholt A and JS King. Coordinated Ras and Rac activity shapes macropinocytic cups and enables phagocytosis of geometrically diverse bacteria. Current Biology, 2020. 30, 2912–2926.

This work demonstrates a new mechanism by which Ras and Rac activity is coordinated to drive macropinocytic and phagocytic cup formation. This suggests a new model for how this occurs in 3D and described the first gene specifically required for the engulfment of complex shapes.

Vines JH and JS King†. The endocytic pathways of Dictyostelium discoideum. Int J Dev Biol. 2019. 63:461-471

Buckley CM, Pots H, Guehu A, Phillips BA, Gilsbach B, Nikolaev A, Soldati T, Parnell AJ, Kortholt, A and King JS. Co-ordinated Ras and Rac activity shapes macropinocytic cups and enables phagocytosis of geometrically diverse bacteria. BioRxiv. 2019, 763748

Itabangi H, Sephton-Clark PCS, Zhou X, Insua I, Probert M, Correia J, Moynihan P, Gebremariam T, Gu Y, Lin L, Ibrahin AS, Brown GS, King JS, Trillo FF, Ballou ER and Voelz K. A bacterial endocymbiont enables fungal immune evasion during fata murcomycete infection. BioRxiv. 2019, 582607

D Sharma, G Otto, E Warren, P Beesley, JS King and RSB Williams. Gamma secretase orthologues are required for lysosomal activity and autophagic degradation in Dictyostelium discoideum, independent of Presenillin proteolytic function. Autophagy, 2019. 15:8, 1407-1418.

CM Buckley, VL Heath, A Gueho, C Bosmani, P Knobloch, P Sikakana, N Personnic, SK Dove, RH Michell, R Meier, H Hilbi, T Soldati, RH Insall†and JS King†. PIKfyve/Fab1 is required for efficient V-ATPase and hydrolase delivery to phagosomes, phagosomal killing, and restriction of Legionella infection. PloS Pathogens. 15 (2), e1007551

This was the first mechanistic study of the role of PIKfyve during phagosome maturation. We show this enzyme regulates phospholipid signalling on the phagosome membrane after engulfment and that this is essential for proper processing. We demonstrate this is critical for phagocytes to kill pathogens and protect themselves from infection.

J Swanson and JS King. The breadth of macropinocytosis research. Phil. Trans. R Soc. B. 2018. 374 (1765), 20180146

JS King†and RR Kay†. The origins and evolution of macropinocytosis. Phil. Trans. R Soc. B. 2019. 374 (1765), 20180158

AT Lopez-Jimenez, EC Munoz, F Leuba, L Gerstenmaier, M Hagedorn, JS King and T Soldati. ESCRT and autophagy cooperate to repair ESX-1-dependent damage to the Mycobacterium-containing vacuole. PLoS Pathogens.2018 Dec 31;14(12):e1007501.

RA Watkins, A Andrews, C Wynn, C Barisch, JS King†and SA Johnston† .Cryptococcus neoformans escape from Dictyostelium amoeba by both WASH-mediated constitutive exocytosis and vomocytosis. Frontiers in Cellular and Infection Microbiology. 2018 doi: 10.3389/fcimb.2018.00108

It is proposed that environmental interactions drive the evolution of fungal virulence.  We therefore investigated how an opportunistic fungal pathogen evades Dictyostelium. We show that the mechanism used by Cryptococcus to escape human macrophages is evolutionarily conserved, but is not the major route used in amoebae, challenging this hypothesis.

RA Watkins, JS King†, SA Johnston†. Nutritional Requirements and Their Importance for Virulence of Pathogenic Cryptococcus Species. Microorganisms 5 (4), 65

Dominguez-Martin E, Cardenal-Munoz E, King JS, Soldati T, Coria R and Escalante R. Methods to Monitor and Quantify Autophagy in the Social Amoeba Dictyostelium discoideum. Cells 2017 6, 18; doi:10.3390/cells6030018

Buckley CM and King JS†. Drinking problems: Mechanisms of macropinosome formation and maturation. FEBS Journal 2017 May 24. doi: 10.1111/febs.14115.

Cardenal-Muñoz E, Arafah S, López-Jiménez AT,Kicka S, Falaise A, Bach F, King JS, Hagedorn M, Soldati T. Mycobacterium marinum antagonistically induces an autophagic response while repressing the autophagic flux in a TORC1- and ESX-1-dependent manner. PLoS Pathogens. 13(4): e1006344.

Thomason P, King JS, Insall RH. Mroh1, a lysosomal regulator localised by WASH-generated actin. J. Cell Sci. May 15;130(10):1785-1795

Mesquita A, Cardenal-Muñoz E, Dominguez E, Muñoz-Braceras S, Nuñez-Corcuera B, Phillips BA, Tábara LC, Xiong Q, Coria R, Eichinger L, Golstein P, King JS, Soldati T, Vincent O, Escalante R. Autophagy in Dictyostelium: Mechanisms, regulation and disease in a simple biomedical model. Autophagy. 2016 Oct 7:1-17.

Buckley CM*, Gopaldass N*, Bosmani C, Johnston SA, Soldati T, Insall RH†, King JS†. WASH drives early recycling from macropinosomes and phagosomes to maintain surface phagocytic receptors. Proceedings of the National Academy of Sciences, USA. 2016 Oct 4;113(40):E5906-E5915

Here, we provide direct evidence for the physiological importance of membrane turnover by macropinocytosis, and uncovered the mechanism by which internalized proteins are recycled by the activity of the WASH complex.

Klionsky DJ. et al. Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy (3rd Edition) Autophagy. 2016;12(1):1-222.

Gerstenmaier L, Pilla R, Herrmann L, Herrmann H, Prado M, Villafano GJ, Kolonko M, Reimer R, Soldati T, King JS, Hagedorn M. The autophagic machinery ensures nonlytic transmission of mycobacteria. Proceedings of the National Academy of Sciences USA, 2015. 112 (7), E687-E692

Urushihara H, Kuwayama H, Fukuhara K, Itoh T, Kagoshima H, Shin-I T, Toyoda A, Ohishi K, Taniguchi T, Noguchi H, Kuroki Y, Hata T, Uchi K, Mohri K, King JS, Insall RH, Kohara Y, Fujiyama A. Comparative genome and transcriptome analyses of the social amoeba Acytostelium subglobosum that accomplishes multicellular development without germ-soma differentiation. BMC Genomics 2015. 16 (1) 80.

Calvo-Garrido J*, King JS*, Munoz-Braceras S, Escalante R. Vmp1 regulates PtdIns3P signaling during autophagosome formation in Dictyostelium discoideum. Traffic 2014. 15, 1235-1246.

King JS†, Gueho A, Hagedorn M, Gopaldass N, Leuba F, Soldati T and Insall RS. WASH is required for lysosomal recycling and efficient autophagic and phagocytic digestion. Mol. Biol. Cell. 2013. 24 (17), 2714-2726

We identified an additional role for the actin-regulator WASH in maintaining lysosomal activity by recycling and delivery of hydrolases to newly-formed autophagosomes and phagosomes.

Davidson AJ*, King, JS*†, RH Insall. The use of streptavidin conjugates as immunoblot loading controls and mitochondrial markers for use with Dictyostelium discoideum. BioTechniques 2013. 55, 39-41

Park L, Thomason PS, Zech T, King JS, Ura S, Carnell M, Machesky LM and Insall RH. Cyclical action of the WASH complex: FAM21 and capping protein drive WASH recycling, not initial recruitment.  Dev. Cell. 2013 Jan 28;24(2):169-81

King, JS†. Mechanical stress meets autophagy: potential implications for physiology and pathology. Trends Mol. Med. 2012. 18(10) pp. 583 – 588

Veltman DM, King JS, Machesky LM and Insall RH. SCAR knockouts in Dictyostelium: WASP assumes SCAR’s position and upstream regulators in pseudopods. J. Cell Biol. 2012; 198(4): 501-508

King JS†. Autophagy across the eukaryotes: Is S. cerevisiae the odd one out?.  Autophagy. 2012 Jul 1;8(7)

Klionsky DJ. et al. Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy, 2012 Apr 1;8(4).

2011

King JS†, Veltman DM and Insall RH. The induction of autophagy by mechanical stress. Autophagy. 2011 Dec 1;7(12).

King JS†, Veltman DM and Insall RH. The induction of autophagy by mechanical stress. Autophagy. 2011 Dec 1;7(12).

King JS†, Veltman DM, Georgiou M, Baum B, Insall RH. SCAR/WAVE is activated at mitosis and drives myosin-independent cytokinesis. J. Cell Sci. 2010 Jul 1;123(Pt 13): 2246-55.

We showed how the cell migration machinery is activated at the end of mitosis to ensure proper division in Dictyostelium cells with a conserved role within metazoan tissue using the Drosophila model.

King JS, Keim M, Teo R, Weening KE, Kapur M, McQuillan K, Ryves J, Rogers B, Dalton E, Williams RSB and Harwood AJ. Genetic control of lithium sensitivity and regulation of inositol biosynthetic genes. PLOS One, 2010 Jun 17;5(6):e11151.

Teo R, King J, Dalton E, Ryves J, Williams RS, Harwood AJ. PtdIns (3, 4, 5) P3 and inositol depletion as a cellular target of mood stabilizers. Biochem Soc Trans. 2009 Oct;37(Pt 5):1110-4.

King JS†, Insall RH. Chemotaxis: finding the way forward with Dictyostelium. Trends Cell Biol. 2009 Oct;19(10):523-30.

King JS*, Teo R*, Ryves J, Reddy JV, Peters O, Orabi B, Hoeller O, Williams RS, Harwood AJ. The mood stabiliser lithium suppresses PIP3 signalling in Dictyostelium and human cells. Dis Model Mech. 2009 May-Jun;2(5-6):306-12.

Despite widespread use, the mechanism of action of mood stabilising drug lithium is unclear. We showed that it has the conserved effect of blocking phosphoinositide signalling, and PIP3-driven cell migration, potentially explaining how it works clinically.

Cho J, King JS, Qian X, Harwood AJ, Shears SB. Dephosphorylation of 2, 3-bisphosphoglycerate by MIPP expands the regulatory capacity of the Rapoport–Luebering glycolytic shunt. Proc Natl Acad Sci U S A. 2008 Apr 22;105(16):5998-6003.

Kortholt A*, King JS*, Keizer-Gunnink I, Harwood AJ, Van Haastert PJ. Phospholipase C regulation of phosphatidylinositol 3, 4, 5-trisphosphate-mediated chemotaxis. Mol Biol Cell. 2007 Dec;18(12):4772-9.

We identified a new regulatory loop, where, contrary to expectation, phosphoinositide signalling (PIP3) is negatively regulated by phospholipase C. We then show physiological significance, in allowing cells to accurately migrate over a wide range of chemoattractant concentrations.

Zaki M*, King J*, Futterer K, Insall RH. Replacement of the essential Dictyostelium Arp2 gene by its Entamoeba homologue using parasexual genetics. BMC Genet. 2007 Jun 6;8:28.

King J and Insall R. Parasexual genetics using axenic cells. Methods Mol Biol. 2006;346:125-35.

King J and Insall RH. Parasexual genetics of Dictyostelium gene disruptions: identification of a ras pathway using diploids. BMC Genet. 2003 Jul 10;4:12.