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The calmodulin gene in Neurospora crassa is required for normal vegetative growth, ultraviolet survival, and sexual development

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Abstract

We isolated a Neurospora crassa mutant of the calmodulin (cmd) gene using repeat-induced point mutation and studied its phenotypes. The cmd RIP mutant showed a defect in growth, reduced aerial hyphae, decreased carotenoid accumulation, a severe reduction in viability upon ultraviolet (UV) irradiation, and a fertility defect. Moreover, meiotic silencing of the cmd gene resulted in a barren phenotype. In addition, we also performed site-directed mutational analysis of the calcium/calmodulin-dependent kinase-2 (Ca2+/CaMK-2), a target of the CaM protein encoded by the cmd gene. The camk-2 S247A and the camk-2 T267A mutants in a homozygous cross, or in a cross with a Δcamk-2 mutant, displayed an intermediate phenotype, suggesting that serine 247 and threonine 267 phosphorylation sites of the Ca2+/CaMK-2 are essential for full fertility in N. crassa. Therefore, CaM in N. crassa is required for normal vegetative growth, UV survival, and sexual development. Additionally, serine 247 and threonine 267 phosphorylation sites are important for the Ca2+/CaMK-2 function.

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Acknowledgements

The FGSC generously waived charges for the Neurospora strains. The FGSC was supported by NSF Grant BIR-9222772. VL was supported by a Research Fellowship from the Ministry of Human Resource Development (MHRD), Government of India (GoI). We thank Professor Katherine A. Borkovich (Department of Plant Pathology and Microbiology, Institute for Integrative Genome Biology, UC Riverside, CA, USA) for help in generating the N. crassa strains expressing the cmd gene under the P tcu-1 . We acknowledge partial financial supports from the IIT Guwahati, Department of Biotechnology, GoI (to RT; BT/PR3635/BCE/8/892/2012), the Indo-US Science and Technology Forum (to RT, 2013/30; http://www.iusstf.org/), and the National Institutes of Health (to KAB; http://www.nih.gov/).

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  1. Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, Assam, 781 039, India

    Vijya Laxmi & Ranjan Tamuli

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  1. Vijya Laxmi
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  2. Ranjan Tamuli
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Correspondence to Ranjan Tamuli.

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Communicated by Erko Stackebrandt.

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Laxmi, V., Tamuli, R. The calmodulin gene in Neurospora crassa is required for normal vegetative growth, ultraviolet survival, and sexual development. Arch Microbiol 199, 531–542 (2017). https://doi.org/10.1007/s00203-016-1319-0

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