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Development and media regulate alternative splicing of a methyltransferase pre-mRNA in Monascus pilosus

Ming-Yong Zhang1, Tsuyoshi Miyake

  • 1South China Botanical Garden, Chinese Academy of Sciences, 723 Xingkelu, Guangzhou 510650, China. zhangmy@scbg.ac.cn

Journal of Agricultural and Food Chemistry
|April 17, 2009
PubMed

Insights

Alternative splicing of the MpLaeA gene in Monascus pilosus produces two mRNA variants. This splicing, influenced by nutrients and growth stage, may regulate monacolin K synthesis.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Biochemistry

Background:

  • The methyltransferase gene MpLaeA in Monascus pilosus exhibits alternative splicing.
  • Two mRNA variants, d-MpLaeA and l-MpLaeA, arise from splicing in the 5'-untranslated region (5'-UTR).

Purpose of the Study:

  • To investigate the regulation of MpLaeA alternative splicing.
  • To determine the functional significance of MpLaeA splicing variants.
  • To explore the role of MpLaeA alternative splicing in monacolin K biosynthesis.

Main Methods:

  • Identification and characterization of MpLaeA mRNA variants in Monascus pilosus.
  • Analysis of alternative splicing patterns under different growth conditions and nutrient sources.
  • Functional analysis of the d-MpLaeA variant using GFP expression in E. coli.
  • Investigating the effect of MpLaeA down-regulation on monacolin K production using transgenic antisense cDNA.

Main Results:

  • Alternative splicing of MpLaeA occurs in the 5'-UTR and is influenced by fungal growth stage and nutrients.
  • The l-MpLaeA mRNA is constitutively expressed, while glutamate and NaNO3 up-regulate the d-MpLaeA mRNA.
  • The long 5'-UTR of d-MpLaeA inhibits gene expression, suggesting it is an ineffective transcript.
  • Down-regulation of MpLaeA led to reduced monacolin K synthesis.

Conclusions:

  • MpLaeA alternative splicing is regulated by environmental factors and growth stages.
  • The d-MpLaeA mRNA variant, due to its long 5'-UTR, appears to be a non-functional transcript.
  • Alternative splicing of MpLaeA likely plays a regulatory role in the biosynthesis of monacolin K.

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