Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
8470935 | Fungal Genetics and Biology | 2013 | 11 Pages |
Abstract
In the n-alkane assimilating yeast Yarrowia lipolytica, the expression of ALK1, encoding a cytochrome P450 that catalyzes terminal mono-oxygenation of n-alkanes, is induced by n-alkanes. The transcription of ALK1 is regulated by a heterocomplex that comprises the basic helix-loop-helix transcription activators, Yas1p and Yas2p, and binds to alkane-responsive element 1 (ARE1) in the ALK1 promoter. An Opi1 family transcription repressor, Yas3p, represses transcription by binding to Yas2p. Yas3p localizes in the nucleus when Y. lipolytica is grown on glucose but localizes to the endoplasmic reticulum (ER) upon the addition of n-alkanes. In this study, we showed that recombinant Yas3p binds to the acidic phospholipids, phosphatidic acid (PA) and phosphoinositides (PIPs), in vitro. The ARE1-mediated transcription was enhanced in vivo in mutants defective in an ortholog of the Saccharomyces cerevisiae gene PAH1, encoding PA phosphatase, and in an ortholog of SAC1, encoding PIP phosphatase in the ER. Truncation mutation analyses for Yas3p revealed two regions that bound to PA and PIPs. These results suggest that the interaction with acidic phospholipids is important for the n-alkane-induced association of Yas3p with the ER membrane.
Keywords
NLSRLM-RACEn-alkaneGSTbHLHDTTeGFPIPTGYarrowia lipolyticaORFP450NaA2D-TLCFFATRNA ligase-mediated rapid amplification of cDNA ends1-naphthaleneacetic acidPhosphatidylinositol 4-phosphatePI(3,4,5)P3PI(4)PPI(4,5)P2basic helix-loop-helixphosphatidic acidSDS-PAGEisopropyl β-D-1-thiogalactopyranosidetwo phenylalanines in an acidic tractdiacylglycerolDiacylglycerol pyrophosphatedithiothreitolDAGTranscriptional repressorCytochrome P450nuclear localization signalendoplasmic reticulumphosphatidylinositolphosphatidylcholinephosphatidylethanolaminePhosphatidylserinephosphatidylinositol 3,4,5-trisphosphatephosphatidylinositol 4,5-bisphosphatePhosphoinositidePhospholipidopen reading framepolymerase chain reactionPCRenhanced green fluorescent proteinPiPTwo-dimensional thin layer chromatographyAIDglutathione S-transferase
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Authors
Satoshi Kobayashi, Kiyoshi Hirakawa, Hiroyuki Horiuchi, Ryouichi Fukuda, Akinori Ohta,