• 제목/요약/키워드: Fumonisin regulation

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Proteomic Comparison of Gibberella moniliformis in Limited-Nitrogen (Fumonisin-Inducing) and Excess-Nitrogen (Fumonisin-Repressing) Conditions

  • Choi, Yoon-E;Butchko, Robert A.E.;Shim, Won-Bo
    • Journal of Microbiology and Biotechnology
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    • 제22권6호
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    • pp.780-787
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    • 2012
  • The maize pathogen Gibberella moniliformis produces fumonisins, a group of mycotoxins associated with several disorders in animals and humans, including cancer. The current focus of our research is to understand the regulatory mechanisms involved in fumonisin biosynthesis. In this study, we employed a proteomics approach to identify novel genes involved in the fumonisin biosynthesis under nitrogen stress. The combination of genome sequence, mutant strains, EST database, microarrays, and proteomics offers an opportunity to advance our understanding of this process. We investigated the response of the G. moniliformis proteome in limited nitrogen (N0, fumonisin-inducing) and excess nitrogen (N+, fumonisin-repressing) conditions by one- and two-dimensional electrophoresis. We selected 11 differentially expressed proteins, six from limited nitrogen conditions and five from excess nitrogen conditions, and determined the sequences by peptide mass fingerprinting and MS/MS spectrophotometry. Subsequently, we identified the EST sequences corresponding to the proteins and studied their expression profiles in different culture conditions. Through the comparative analysis of gene and protein expression data, we identified three candidate genes for functional analysis and our results provided valuable clues regarding the regulatory mechanisms of fumonisin biosynthesis.

Regulation of Fumonisin Biosynthesis in Fusarium verticillioides-Maize System

  • Sagaram Uma Shankar;Kolomiets Mike;Shim Won-Bo
    • The Plant Pathology Journal
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    • 제22권3호
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    • pp.203-210
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    • 2006
  • Fumonisins are a group of mycotoxins produced by a pathogen Fusarium verticillioides in infected maize kernels. Consumption of fumonisin-contaminated maize has been implicated in a number of animal and human illnesses, including esophageal cancer and neural tube defects. Since the initial discovery, chemistry, toxicology, and biology of fumonisins as well as the maize-Fusarium pathosystem have been extensively studied. Furthermore, in the past decade, significant progress has been made in terms of understanding the molecular biology of toxin biosynthetic genes. However, there is a critical gap in our understanding of the regulatory mechanisms involved in fumonisin biosynthesis. Here, we review and discuss our current knowledge about the molecular mechanisms by which fumonisin biosynthesis is regulated in F. verticillioides. In addition, we discuss the impact of maize kernel environment, particularly sugar and lipid molecules, on fumonisin biosynthesis.

Identification of Genes Associated with Fumonisin Biosynthesis in Fusarium verticillioides via Proteomics and Quantitative Real-Time PCR

  • Choi, Yoon-E.;Shim, Won-Bo
    • Journal of Microbiology and Biotechnology
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    • 제18권4호
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    • pp.648-657
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    • 2008
  • In this study, we used functional genomic strategies, proteomics and quantitative real-time (qRT)-PCR, to advance our understanding of genes associated with fumonisin production in the fungus Fusarium verticillioides. Earlier studies have demonstrated that deletion of the FCC1 gene, which encodes a C-type cyclin, leads to a drastic reduction in fumonisin production and conidiation in the mutant strain (FT536). The premise of our research was that comparative analysis of F. verticillioides wild-type and FT536 proteomes will reveal putative proteins, and ultimately corresponding genes, that are important for fumonisin biosynthesis. We isolated proteins that were significantly upregulated in either the wild type or FT536 via two-dimensional polyacrylamide gel electrophoresis, and subsequently obtained sequences by mass spectrometry. Homologs of identified proteins, e.g., carboxypeptidase, laccase, and nitrogen metabolite repression protein, are known to have functions involved in fungal secondary metabolism and development. We also identified gene sequences corresponding to the selected proteins and investigated their transcriptional profiles via quantitative real-time (qRT)-PCR in order to identify genes that show concomitant expression patterns during fumonisin biosynthesis. These genes can be selected as targets for functional analysis to further verify their roles in $FB_1$ biosynthesis.

Geranyllinalool에 의한 LLC-PK1 세포내 스핑고지질 생합성 억제 (Inhibition of de Novo Sphingolipid Biosynthesis by Geranyllinalool in $LLC-PK_1$ Cells)

  • 조양혁;이용문
    • 약학회지
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    • 제43권1호
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    • pp.61-67
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    • 1999
  • Geranyllinalool, a polyisoprenoid compound, was found to block the early biosynthetic pathway of sphingolipids in LLC-PKl cells. Sphinganine, an intermediate in sphingolipid biosynthetic pathway, was abruptly accumulated in LLC-PKl cells at $2{\;}{\mu}M$ of fumonisin B1(FB1), a specific inhibitor of sphinganine N-acyltransferase, for 24 hr. Geranyllinalool lowered the $B_1(FB_1)$, a specific inhibitor of sphinganine N-acyltransferase, for 24 hr. Geranyllinalool lowered th FB1 and $50{\;}\mu$M geranyllinalool. l-Cy-closerine, an inhibitor of serine-palmitoyl transferase, was used as a positive control to evaluate the inhibitory effect of geranyllinalool. These results suggest that geranyllinalool may inhibit the serine-palmitoyl transferase, the first enzyme in de novo sphingolipid biosynthesis, resulting in the altered regulation of sphingolipid metabolism.

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2009년산 옥수수와 벼에서의 Fusarium 곰팡이독소 자연발생량 조사 (Natural Occurrence of Fusarium Mycotoxins in Field-collected Maize and Rice in Korea in 2009)

  • 이승호;손승완;남영주;신진영;이수형;김미자;윤종철;류재기;이데레사
    • 식물병연구
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    • 제16권3호
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    • pp.306-311
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    • 2010
  • 2009년 국내산 옥수수 19점과 벼 32점을 대상으로 Fusarium 오염 및 Fusarium 독소 오염을 조사하였다. 옥수수와 벼 시료의 Fusarium 오염률은 각각 54.9%와 8.2%로 확인되었으며, 종 특이 primer를 이용한 PCR 증폭결과 옥수수시료에서 분리된 506균주 중 58균주의 F.graminearum 추정균주(11.5%)와 354균주의 F. verticillioides 추정균주(70.0%)가 동정되었다. 또한 벼의 경우, 분리된 315균주 중 276균주(87.8%)가 F. graminearum으로 추정 되었으며, F. verticillioides 추정균주는 검출되지 않았다. LC 및 LC-MS를 이용한 Fusarium 독소(DON, NIV, ZEA, FB)의 자연발생량 조사 결과, DON과 ZEA이 각각 2개의 옥수수 시료에서만 기준치 이상 검출되었다. FB는 대부분의 옥수수 시료와 한 개의 벼 시료에서 검출 되었으나 모두 기준치 이하였다. 따라서 본 연구에서 사용된 2009년산 옥수수와 벼의 곰팡이독소 오염수준은 대부분 기준치 이하로 심각하지 않았다.

Links between accelerated replicative cellular senescence and down-regulation of SPHK1 transcription

  • Kim, Min Kyung;Lee, Wooseong;Yoon, Gang-Ho;Chang, Eun-Ju;Choi, Sun-Cheol;Kim, Seong Who
    • BMB Reports
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    • 제52권3호
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    • pp.220-225
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    • 2019
  • We have identified a mechanism to diminish the proliferative capacity of cells during cell expansion using human adipose-derived stromal cells (hAD-SCs) as a model of replicative senescence. hAD-SCs of high-passage numbers exhibited a reduced proliferative capacity with accelerated cellular senescence. Levels of key bioactive sphingolipids were significantly increased in these senescent hAD-SCs. Notably, the transcription of sphingosine kinase 1 (SPHK1) was down-regulated in hAD-SCs at high-passage numbers. SPHK1 knockdown as well as inhibition of its enzymatic activity impeded the proliferation of hAD-SCs, with concomitant induction of cellular senescence and accumulation of sphingolipids, as seen in high-passage cells. SPHK1 knockdown-accelerated cellular senescence was attenuated by co-treatment with sphingosine-1-phosphate and an inhibitor of ceramide synthesis, fumonisin $B_1$, but not by treatment with either one alone. Together, these results suggest that transcriptional down-regulation of SPHK1 is a critical inducer of altered sphingolipid profiles and enhances replicative senescence during multiple rounds of cell division.