Journal of Microbiology

Keon-Sang Chae

6 Articles

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; 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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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.

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A Novel Rapid Fungal Promoter Analysis System Using the Phosphopantetheinyl Transferase Gene,npgA, inAspergillus nidulans
Ha-Yeon Song, Dahye Choi, Dong-Min Han, Dae-Hyuk Kim, Jung-Mi Kim
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Both a PKS and a PPTase are involved in melanin biosynthesis and regulation of Aureobasidium melanogenum XJ5-1 isolated from the Taklimakan desert
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Identification of a Polyketide Synthase Gene in the Synthesis of Phleichrome of the Phytopathogenic Fungus Cladosporium phlei
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The MpkB MAP Kinase Plays a Role in Post-karyogamy Processes as well as in Hyphal Anastomosis During Sexual Development in Aspergillus nidulans: Sang-Cheol Jun , Sei-Jin Lee , Hyun-Joo Park , Ji-Young Kang , Young-Eun Leem , Tae-Ho Yang , Mi-Hee Chang , Jung-Mi Kim , Seung-Hwan Jang , Hwan-Gyu Kim , Dong-Min Han , Keon-Sang Chae , Kwang-Yeop Jahng; J. Microbiol. 2011;49(3):418-430. Published online June 30, 2011; DOI: https://doi.org/10.1007/s12275-011-0193-3

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Abstract PDF

Two genes encoding MAP kinase homologs, designated as mpkB and mpkC, were isolated from Aspergillus nidulans by PCR with degenerate primers. Deletion and over-expression mutants of mpkC showed no detectable phenotypes under any external stress tested. Deletion of mpkB caused pleiotropic phenotypes including a failure in forming cleistothecia under any induction conditions for sexual development, increased Hülle cell production, slow hyphal growth and aberrant conidiophore morphology. Over-expression of mpkB led to increased cleistothecium production. While the transcripts of mpkB and mpkC were constitutively synthesized through the entire life cycle, their size and amount differed with developmental stages. An outcross test using fluorescent protein reporters showed that the mpkB deletion mutant could not form heterokaryons with wild type. Protoplast fusion experiments showed that the fusant of the mpkB mutant with wild type could undergo normal sexual development. However, heterokaryotic mycelia that were produced from a fusant between two mpkB deletion mutants could not form cleistothecia, although they did appear to form diploid nuclei. These results suggest that the MpkB MAP kinase is required for some post-karyogamy process as well as at the hyphal anastomosis stage to accomplish sexual development successfully.

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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
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Predicting the Chemical Composition and Structure of Aspergillus nidulans Hyphal Wall Surface by Atomic Force Microscopy: Hyun-uk Lee , Jong Bae Park , Haeseong Lee , Keon-Sang Chae , Dong-Min Han , Kwang-Yeop Jahng; J. Microbiol. 2010;48(2):243-248. Published online May 1, 2010; DOI: https://doi.org/10.1007/s12275-010-8094-4

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Abstract PDF

In fungi, cell wall plays an important role in growth and development. Major macromolecular constituents of the aspergilli cell wall are glucan, chitin, and protein. We examined the chemical composition and structure of the Aspergillus nidulans hyphal wall surface by an atomic force microscope (AFM). To determine the composition of the cell wall surface, the adhesion forces of commercially available β-glucan, chitin, and various proteins were compared to those of corresponding fractions prepared from the hyphal wall. In both setups, the adhesion forces of β-glucan, chitin, and protein were 25-50, 1000-3000, and 125-300 nN, respectively. Adhesion force analysis demonstrated that the cell surface of the apical tip region might contain primarily chitin and β-glucan and relatively a little protein. This analysis also showed the chemical composition of the hyphal surface of the mid-region would be different from that of the apical region. Morphological images obtained by the tapping mode of AFM revealed that the hyphal tip surface has moderate roughness.

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Effect of cell wall integrity stress and RlmA transcription factor on asexual development and autolysis in Aspergillus nidulans
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Regulation of nsdD Expression in Aspergillus nidulans: Kap Hoon Han , Kyu-Yong Han , Min-Su Kim , Dong-Beom Lee , Jong-Hak Kim , Suhn-Kee Chae , Keon-Sang Chae , Dong-Min Han; J. Microbiol. 2003;41(3):259-261.

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Abstract PDF: The nsdD gene has been predicted to encode a GATA type transcription factor with the type IVb zinc finger DNA binding domain functions in activating sexual development of A. nidulans. In several allelic mutants of nsdD producing truncated NsdD polypeptides lacking the C-terminal zinc finger, the transcription level of nsdD gene was greatly increased. Also in an over-expressed mutant, the transcription under its own promoter was reduced. These results suggest that the expression of nsdD is negatively autoregulated. When the nsdD gene was over-expressed, cleistothecia were formed in excess amounts even in the presence of 0.6M KCl that inhibited sexual development of the wild type. Northern blot analysis revealed that the expression of nsdD was repressed by 0.6M KCl. These results strongly suggest that the inhibition of sexual development by salts was carried out via the nsdD involved regulatory network.

Ultrastructure of the Cell Wall of a Null Pigmentation Mutant, npgA1, in Aspergillus nidulans: Yun Shin Chung , § , Jung-Mi Kim , Dong-Min Han , Keon-Sang Chae , Kwang-Yeop Jahng; J. Microbiol. 2003;41(3):224-231.

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Abstract PDF: The null pigmentation mutant (npgA1) of Aspergillus nidulans was previously characterized by its production of no pigment at any stage of its life cycle, its reduction in hyphal branching, and its delay in the asexual spore development. The chemical composition of the cell wall was also altered in npgA1 mutants that became more sensitive to Novozyme 234_TM, which is possibly due to a structural defect in the cell wall. To investigate the effects of the cell wall structure on these pleiomorphic phenomena, we examined the ultrastructure of the cell wall in the npgA1 mutant (WX17). Scanning electron micrographs (SEM) showed that after being cultured for six days, the outermost layer of the conidial wall of WX17 peeled off. Although this phenotype suggested that the cell wall structure in WX17 may be modified, examination using TEM of the fine structure of cross-sectioned hyphal wall of WX17 did not show any differences from that of FGSC4. However, staining for carbohydrates of wall layers showed that the electron-translucent layer of the cell wall was missing in WX17. In addition, the outermost layer H1 of the hyphal wall was also absent in WX17. The ultrastructural observation and cytochemical analysis of cell walls suggested that the pigmentation defect in WX17 may be attributed to the lack of a layer in the cell wall.

Promotion of Asexual Development and Inhibition of Sexual Development of Aspergillus nidulans by Short-Chain Primary Amines: Myung Hoon Song , Kuppusamy Selvam , Chang-Jun Choi , Kwang-Yeop Jahng , Dong-Min Han , Keon-Sang Chae; J. Microbiol. 2002;40(3):230-233.

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Abstract PDF: Effects of short-chain primary amines on Aspergillus nidulans development were analyzed. Propylamine induced asexual development and inhibited sexual development. Even on medium containing lactose as the sole carbon source, on which little conidial heads are formed and sexual structures are formed preferentially, or when sexual development was induced, propylamine induced asexual development and inhibited sexual development. These effects of propylamine seemed to be due to accumulation of mRNA of the brlA gene, which has been identified as a positive regulator of asexual development, and due to the reduction of the veA mRNA level. The veA gene has been identified as an activator of sexual development and also as an inhibitor of asexual development. Other primary amines, methylamine and ethylamine, showed identical effects on development where short-chain primary amine also promoted asexual development and inhibited sexual development.

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