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Rapamycin induces morphological and physiological changes without increase in lipid content in Ustilago maydis

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Abstract

The evolutionarily conserved serine/threonine kinase TOR recruits different subunits to assemble the Target of Rapamycin Complex 1 (TORC1), which is inhibited by rapamycin and regulates ribosome biogenesis, autophagy, and lipid metabolism by regulating the expression of lipogenic genes. In addition, TORC1 participates in the cell cycle, increasing the length of the G2 phase. In the present work, we investigated the effect of rapamycin on cell growth, cell morphology and neutral lipid metabolism in the phytopathogenic fungus Ustilago maydis. Inhibition of TORC1 by rapamycin induced the formation of septa that separate the nuclei that were formed after mitosis. Regarding neutral lipid metabolism, a higher accumulation of triacylglycerols was not detected, but the cells did contain large lipid bodies, which suggests that small lipid bodies became fused into big lipid droplets. Vacuoles showed a similar behavior as the lipid bodies, and double labeling with Blue-CMAC and BODIPY indicates that vacuoles and lipid bodies were independent organelles. The results suggest that TORC1 has a role in cell morphology, lipid metabolism, and vacuolar physiology in U. maydis.

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Acknowledgements

This research was supported by the Universidad Nacional Autónoma de México, Programa de Apoyo a Proyectos de Investigación Tecnológica PAPIIT IN222117; Consejo Nacional de Ciencia y Tecnología CONACYT 254904-JPP and 256520-GGS; Instituto Politécnico Nacional-Secretaria de Investigación y Posgrado IPN-SIP-20180625. We thank Universidad Nacional Autónoma de México, Programa de Becas Posdoctorales en la UNAM POSDOC-Dirección General de Asuntos del Personal Académico DGAPA–UNAM for the fellowship granted to Lucero Romero–Aguilar. We thank QFB. Oscar Ivan Luqueño–Bocardo for technical assistance, and Dr. Jorge Guevara Fonseca for the equipment provided.

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Authors and Affiliations

  1. Departamento de Bioquímica, Facultad de Medicina, Universidad Nacional Autónoma de México, Avenida Universidad 3000, Cd. Universitaria, Coyoacán, 04510, Mexico City, Mexico

    Lucero Romero-Aguilar, Eda P. Tenorio, Genaro Matus-Ortega, Oscar Flores-Herrera & Juan Pablo Pardo

  2. Departamento de Microbiología, Instituto Politécnico Nacional, Escuela Nacional de Ciencias Biológicas, Prolongación de Carpio Y Plan de Ayala S/N, Miguel Hidalgo, 11340, Mexico City, Mexico

    Guadalupe Guerra-Sánchez

  3. Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Avenida Universidad 3000, Cd. Universitaria, Coyoacán, 04510, Mexico City, Mexico

    Miguel Tapia-Rodriguez

  4. Departamento de Biología Celular, Facultad de Ciencias, Universidad Nacional Autónoma de México, Avenida Universidad 3000, Cd. Universitaria, Coyoacán, 04510, Mexico City, Mexico

    James González

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  1. Lucero Romero-Aguilar
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Contributions

LRA conceptualized the idea, performed microbiological experiments, FACS analysis, wrote and reviewed the manuscript. JPP conceptualized and coordinated the research, analyzed bioinformatics data, wrote the manuscript and provided funding to support the research. GGS analyzed the results, provided funding to support the research, reviewed and corrected the manuscript. GMO performed bioinformatics analysis, microbiological experiments, reviewed and corrected the manuscript. MTR performed confocal microscopy. EPT performed FACS analysis, reviewed and corrected the manuscript. OFH reviewed and corrected the manuscript. JG performed microbiological experiments, reviewed and corrected the manuscript.

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Correspondence to Juan Pablo Pardo.

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Communicated by Erko Stackebrandt.

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Electronic supplementary material

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203_2020_1833_MOESM1_ESM.pdf

Pairwise sequence alignment of Tor amino acid sequences from Saccharomyces cerevisiae and Ustilago maydis. Alignment was performed with Clustal W

203_2020_1833_MOESM2_ESM.pdf

Pairwise sequence alignment of Tor amino acid sequences from Homo sapiens and Ustilago maydis. Alignment was performed with Clustal W

203_2020_1833_MOESM3_ESM.mp4

3D reconstruction of cells labeled with Blue-CMAC and BODIPY. XYZ capture was done accordingly with the Nyquist-Shannon sampling theorem and was reconstructed with NIS Elements v5.0 software. FB2 yeast cells were cultured during 24 hours in minimal medium lacking a nitrogen 674 source.

203_2020_1833_MOESM4_ESM.docx

Components of the Greatwall-Endosulfine, TORC1, and cell cycle pathways, in U. maydis. A BLASTp was carried out using S. pombe amino acid sequences as queries

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Romero-Aguilar, L., Guerra-Sánchez, G., Tenorio, E.P. et al. Rapamycin induces morphological and physiological changes without increase in lipid content in Ustilago maydis. Arch Microbiol 202, 1211–1221 (2020). https://doi.org/10.1007/s00203-020-01833-y

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