Characterization of NpgA, a 4'-phosphopantetheinyl transferase of Aspergillus nidulans, and evidence of its involvement in fungal growth and formation of conidia and cleistothecia for development

Jung-Mi Kim; Ha-Yeon Song; Hyo-Jin Choi; Kum-Kang So; Dae-Hyuk Kim; Keon-Sang Chae; Dong-Min Han; Kwang-Yeop Jahng

doi:10.1007/s12275-015-4657-8

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Characterization of NpgA, a 4'-phosphopantetheinyl transferase of Aspergillus nidulans, and evidence of its involvement in fungal growth and formation of conidia and cleistothecia for development: Jung-Mi Kim ¹, Ha-Yeon Song ¹, Hyo-Jin Choi ¹, Kum-Kang So ², Dae-Hyuk Kim ², Keon-Sang Chae ², Dong-Min Han ³, Kwang-Yeop Jahng ⁴; Journal of Microbiology 2015;53(1):21-31.
DOI: https://doi.org/10.1007/s12275-015-4657-8
Published online: January 4, 2015

¹Department of Bio-Environmental Chemistry, Wonkwang University, Iksan 570-749, Republic of Korea, ²Department of Molecular Biology, Institute for Molecular Biology and Genetics, Center for Fungal Pathogenesis, Chonbuk National University, Jeonju 561-756, Republic of Korea, ³Division of Biological Sciences, Wonkwang University, Iksan 570-749, Republic of Korea, ⁴Department of Biological Science, Chonbuk National University, Jeonju 561-756, Republic of Korea¹Department of Bio-Environmental Chemistry, Wonkwang University, Iksan 570-749, Republic of Korea, ²Department of Molecular Biology, Institute for Molecular Biology and Genetics, Center for Fungal Pathogenesis, Chonbuk National University, Jeonju 561-756, Republic of Korea, ³Division of Biological Sciences, Wonkwang University, Iksan 570-749, Republic of Korea, ⁴Department of Biological Science, Chonbuk National University, Jeonju 561-756, Republic of Korea

Corresponding author: Kwang-Yeop Jahng , Tel: +82-63-270-3358,

Received: 14 November 2014 • Revised: 15 December 2014 • Accepted: 15 December 2014

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Abstract

The null pigmentation mutant (npgA1) in Aspergillus nidulans
results
in a phenotype with colorless organs, decreased branching growth, delayed of asexual spore development, and aberrant cell wall structure. The npgA gene was isolated from A. nidulans to investigate these pleiomorphic phenomena of npgA1 mutant. Sequencing analysis of the complementing gene indicated that it contained a 4􍿁-phosphopantetheinyl transferase (PPTase) superfamily domain. Enzymatic assay of the PPTase, encoded by the npgA gene, was implemented in vivo and in vitro. Loss-of-function of LYS5, which encoded a PPTase in Saccharomyces cerevisiae, was functionally complemented by NpgA, and Escherichia coli-derived NpgA revealed phosphopantetheinylation activity with the elaboration of 3􍿁5􍿁-ADP. Deletion of the npgA gene caused perfectly a lethal phenotype and the absence of asexual/sexual sporulation and secondary metabolites such as pigments in A. nidulans. However, a cross feeding effect with A. nidulans wild type allowed recovery from deletion defects, and phased-culture filtrate from the wild type were used to verify that the npgA gene was essential for formation of metabolites needed for development as well as growth. In addition, forced expression of npgA promoted the formation of conidia and cleistothecia as well as growth. These results indicate that the npgA gene is involved in the phosphopantetheinylation required for primary biological processes such as growth, asexual/sexual development, and the synthesis of secondary metabolites in A. nidulans.

Keywords: Aspergillus nidulans;npgA;phosphopantetheinyl transferase;metabolite;development

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Characterization of NpgA, a 4'-phosphopantetheinyl transferase of Aspergillus nidulans, and evidence of its involvement in fungal growth and formation of conidia and cleistothecia for development

J. Microbiol. 2015;53(1):21-31. Published online January 4, 2015

DOI: https://doi.org/10.1007/s12275-015-4657-8

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