Niemann-Pick Type C Proteins Are Required for Sterol Transport and Appressorium-Mediated Plant Penetration of

Overview

Journal mBio

Publisher American Society for Microbiology

Specialty Microbiology

Date 2022 Sep 26

PMID 36154185

Authors

Sayo Kodama

Naoki Kajikawa

Fumi Fukada

Yasuyuki Kubo

Affiliations

Soon will be listed here.

Abstract

Many biotrophic and hemibiotrophic fungal pathogens use appressoria to directly penetrate the host plant surface. In the cucumber anthracnose fungus , differentiation of appressoria requires a proper G/S cell cycle progression, regulated by the GTPase-activating protein complex CoBub2-CoBfa1 and its downstream GTPase CoTem1. To explore the mechanisms by which the CoTem1 cascade regulates plant infection, we screened for CoTem1 interaction factors and identified a Niemann-Pick type C2 homolog (CoNpc2). Niemann-Pick type C proteins NPC1 and NPC2 are sterol-binding proteins required for sterol export from lysosomes (vacuoles) in humans and yeasts. We showed that CoNpc2 colocalized with CoNpc1 in late endosomes and vacuoles and that disruption of its gene resulted in aberrant sterol accumulation in vacuoles and loss of sterol membrane localization, indicating that NPC proteins are engaged in sterol transport in . For appressorium infection, sterol transport and proper distribution mediated by CoNpc1 and CoNpc2 are critical for membrane integrity and membrane curvature with actin assembly, leading to penetration peg emergence and appressorial cone formation. Our results revealed a novel mechanism by which NPC proteins regulate appressorium-mediated plant infection. Fungal morphogenesis requires accurate cell cycle progression. Two-component GTPase-activating protein (GAP) CoBub2-CoBfa1 interacts with downstream GTPase CoTem1 and is required for G/S progression to establish plant infection in To understand the pathogenicity related functions of CoTem1 downstream, we identified a Niemann-Pick type C2 homolog (CoNpc2) as a novel physical interaction factor with CoTem1. Whereas NPC proteins (NPC1 and NPC2) are essential for sterol homeostasis in humans and yeasts, their functions in plant invasion by pathogenic fungi have remained unclear. In this study, we show that and play a critical role in intracellular sterol transport and that appropriate sterol distribution is required for membrane integrity and membrane curvature with actin assembly that leads to appressorium-mediated plant penetration and pathogenicity of . Our findings suggest the importance of sterol distribution in fungal morphogenesis during plant infection.

References

Gan P, Ikeda K, Irieda H, Narusaka M, OConnell R, Narusaka Y . Comparative genomic and transcriptomic analyses reveal the hemibiotrophic stage shift of Colletotrichum fungi. New Phytol. 2012; 197(4):1236-1249. DOI: 10.1111/nph.12085. View

Sleat D, Wiseman J, El-Banna M, Price S, Verot L, Shen M . Genetic evidence for nonredundant functional cooperativity between NPC1 and NPC2 in lipid transport. Proc Natl Acad Sci U S A. 2004; 101(16):5886-91. PMC: 395893. DOI: 10.1073/pnas.0308456101. View

Rosenbaum A, Maxfield F . Niemann-Pick type C disease: molecular mechanisms and potential therapeutic approaches. J Neurochem. 2010; 116(5):789-95. PMC: 3008286. DOI: 10.1111/j.1471-4159.2010.06976.x. View

Friederichs J, Ghosh S, Smoyer C, McCroskey S, Miller B, Weaver K . The SUN protein Mps3 is required for spindle pole body insertion into the nuclear membrane and nuclear envelope homeostasis. PLoS Genet. 2011; 7(11):e1002365. PMC: 3219597. DOI: 10.1371/journal.pgen.1002365. View

Bruno K, Morrell J, Hamer J, Staiger C . SEPH, a Cdc7p orthologue from Aspergillus nidulans, functions upstream of actin ring formation during cytokinesis. Mol Microbiol. 2001; 42(1):3-12. DOI: 10.1046/j.1365-2958.2001.02605.x. View

Heilig Y, Schmitt K, Seiler S . Phospho-regulation of the Neurospora crassa septation initiation network. PLoS One. 2013; 8(10):e79464. PMC: 3804505. DOI: 10.1371/journal.pone.0079464. View

Saarikangas J, Zhao H, Pykalainen A, Laurinmaki P, Mattila P, Kinnunen P . Molecular mechanisms of membrane deformation by I-BAR domain proteins. Curr Biol. 2009; 19(2):95-107. DOI: 10.1016/j.cub.2008.12.029. View

Maxfield F, Menon A . Intracellular sterol transport and distribution. Curr Opin Cell Biol. 2006; 18(4):379-85. DOI: 10.1016/j.ceb.2006.06.012. View

Douglas L, Konopka J . Fungal membrane organization: the eisosome concept. Annu Rev Microbiol. 2014; 68:377-93. DOI: 10.1146/annurev-micro-091313-103507. View

10.

Jacquier N, Schneiter R . Mechanisms of sterol uptake and transport in yeast. J Steroid Biochem Mol Biol. 2010; 129(1-2):70-8. DOI: 10.1016/j.jsbmb.2010.11.014. View

11.

Kumar S, Stecher G, Li M, Knyaz C, Tamura K . MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms. Mol Biol Evol. 2018; 35(6):1547-1549. PMC: 5967553. DOI: 10.1093/molbev/msy096. View

12.

Mendgen K, Deising H . Infection structures of fungal plant pathogens - a cytological and physiological evaluation. New Phytol. 2021; 124(2):193-213. DOI: 10.1111/j.1469-8137.1993.tb03809.x. View

13.

Pan X, Roberts P, Chen Y, Kvam E, Shulga N, Huang K . Nucleus-vacuole junctions in Saccharomyces cerevisiae are formed through the direct interaction of Vac8p with Nvj1p. Mol Biol Cell. 2000; 11(7):2445-57. PMC: 14931. DOI: 10.1091/mbc.11.7.2445. View

14.

Berger A, Vanderford T, Gernert K, Nichols J, Faundez V, Corbett A . Saccharomyces cerevisiae Npc2p is a functionally conserved homologue of the human Niemann-Pick disease type C 2 protein, hNPC2. Eukaryot Cell. 2005; 4(11):1851-62. PMC: 1287848. DOI: 10.1128/EC.4.11.1851-1862.2005. View

15.

Simons K, Gerl M . Revitalizing membrane rafts: new tools and insights. Nat Rev Mol Cell Biol. 2010; 11(10):688-99. DOI: 10.1038/nrm2977. View

16.

Tanaka S, Yamada K, Yabumoto K, Fujii S, Huser A, Tsuji G . Saccharomyces cerevisiae SSD1 orthologues are essential for host infection by the ascomycete plant pathogens Colletotrichum lagenarium and Magnaporthe grisea. Mol Microbiol. 2007; 64(5):1332-49. DOI: 10.1111/j.1365-2958.2007.05742.x. View

17.

Simunovic M, Voth G, Callan-Jones A, Bassereau P . When Physics Takes Over: BAR Proteins and Membrane Curvature. Trends Cell Biol. 2015; 25(12):780-792. PMC: 4831700. DOI: 10.1016/j.tcb.2015.09.005. View

18.

Ikonen E . Cellular cholesterol trafficking and compartmentalization. Nat Rev Mol Cell Biol. 2008; 9(2):125-38. DOI: 10.1038/nrm2336. View

19.

Kodedova M, Sychrova H . Changes in the Sterol Composition of the Plasma Membrane Affect Membrane Potential, Salt Tolerance and the Activity of Multidrug Resistance Pumps in Saccharomyces cerevisiae. PLoS One. 2015; 10(9):e0139306. PMC: 4587746. DOI: 10.1371/journal.pone.0139306. View

20.

Gong X, Qian H, Zhou X, Wu J, Wan T, Cao P . Structural Insights into the Niemann-Pick C1 (NPC1)-Mediated Cholesterol Transfer and Ebola Infection. Cell. 2016; 165(6):1467-1478. PMC: 7111323. DOI: 10.1016/j.cell.2016.05.022. View