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Control of morphology and virulence by ADP-ribosylation factors (Arf) in Mucor circinelloides

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Abstract

Mucor circinelloides is a dimorphic fungus used to study cell differentiation that has emerged as a model to characterize mucormycosis. In this work, we identified four ADP-ribosylation factor (Arf)-encoding genes (arf1–arf4) and study their role in the morphogenesis and virulence. Arfs are key regulators of the vesicular trafficking process and are associated with both growth and virulence in fungi. Arf1 and Arf2 share 96% identity and Arf3 and Arf4 share 89% identity, which suggests that the genes arose through gene-duplication events in M. circinelloides. Transcription analysis revealed that certain arf genes are affected by dimorphism of M. circinelloides, such as the arf2 transcript, which was accumulated during yeast development. Therefore, we created knockout mutants of four arf genes to evaluate their function in dimorphism and virulence. We found that both arf1 and arf2 are required for sporulation, but these genes also perform distinct functions; arf2 participates in yeast development, whereas arf1 is involved in aerobic growth. Conversely, arf3 and arf4 play only minor roles during aerobic growth. Moreover, we observed that all single arf-mutant strains are more virulent than the wild-type strain in mouse and nematode models, with the arf3 mutant being most virulent. Lastly, arf1/arf2 and arf3/arf4 double mutations produced heterokaryon strains that did not reach the homokaryotic state, indicating that these genes participate in essential and redundant functions. Overall, this work reveals that Arfs proteins regulate important cellular processes in M. circinelloides such as morphogenesis and virulence, laying the foundation to characterize the molecular networks underlying this regulation.

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Acknowledgements

This work was supported by grants from Coordinación de la Investigación Científica, UMSNH, México (2.6, 2.35), Consejo Nacional de Ciencia y Tecnología, México (CONACYT; 181747, 167071, and 256119), and Fundación Séneca-Agencia de Ciencia y Tecnología de la Región de Murcia, Spain (19339/PI/14). JA P-M, G M-H, MI V-M, and NY R-M were supported by postgraduate fellowships from CONACYT and CP-A by Ministerio de Educación, Cultura y Deporte of Spain (FPU-14/01832).

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Authors and Affiliations

  1. Laboratorio de Diferenciación Celular, Instituto de Investigaciones Químico Biológicas, Universidad Michoacana de San Nicolás de Hidalgo, Ciudad Universitaria, 58030, Morelia, Michoacan, Mexico

    J. Alberto Patiño-Medina, Guadalupe Maldonado-Herrera, Viridiana Alejandre-Castañeda, Nancy Y. Reyes-Mares, Marco I. Valle-Maldonado, Jesus Campos-García, Martha I. Ramírez-Díaz & Victor Meza-Carmen

  2. Departamento de Genética y Microbiología, Facultad de Biologia, Universidad de Murcia, 30100, Murcia, Spain

    Carlos Pérez-Arques & Victoriano Garre

  3. Facultad de Químico Farmacobiología, Universidad Michoacana de San Nicolás de Hidalgo, Morelia, Michoacan, Mexico

    Rafael Ortiz-Alvarado

  4. Laboratorio Estatal de Salud Pública del Estado de Michoacán, 58279, Morelia, Michoacan, Mexico

    Irvin E. Jácome-Galarza

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  1. J. Alberto Patiño-Medina
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  2. Guadalupe Maldonado-Herrera
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  3. Carlos Pérez-Arques
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  4. Viridiana Alejandre-Castañeda
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  5. Nancy Y. Reyes-Mares
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  6. Marco I. Valle-Maldonado
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  7. Jesus Campos-García
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  8. Rafael Ortiz-Alvarado
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  10. Martha I. Ramírez-Díaz
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  11. Victoriano Garre
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  12. Victor Meza-Carmen
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Contributions

JAP-M, GM-H, MIV-M, CP-A, VA-C, NYR-M, RO-A, data acquisition and interpretation; RO-A, IEJ-G, JC-G, MIR-D, VG, reagents, critical analysis, and revision of the article for intellectual content; MIR-D, VG, VM-C, conception and design of the work and data analysis and interpretation; VM-C, drafting of the article.

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Correspondence to Victor Meza-Carmen.

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Communicated by M. Kupiec.

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Patiño-Medina, J.A., Maldonado-Herrera, G., Pérez-Arques, C. et al. Control of morphology and virulence by ADP-ribosylation factors (Arf) in Mucor circinelloides . Curr Genet 64, 853–869 (2018). https://doi.org/10.1007/s00294-017-0798-0

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