Engineering Promoter Architecture in Oleaginous Yeast Yarrowia lipolytica.
Eukaryotic promoters have a complex architecture to control both the strength and timing of gene transcription spanning up to thousands of bases from the initiation site. This complexity makes rational fine-tuning of promoters in fungi difficult to predict; however, this very same complexity enables multiple possible strategies for engineering promoter strength. Here, we studied promoter architecture in the oleaginous yeast, Yarrowia lipolytica. While recent studies have focused on upstream activating sequences, we systematically examined various components common in fungal promoters. Here, we examine several promoter components including upstream activating sequences, proximal promoter sequences, core promoters, and the TATA box in autonomously replicating expression plasmids and integrated into the genome. Our findings show that promoter strength can be fine-tuned through the engineering of the TATA box sequence, core promoter, and upstream activating sequences. Additionally, we identified a previously unreported oleic acid responsive transcription enhancement in the XPR2 upstream activating sequences, which illustrates the complexity of fungal promoters. The promoters engineered here provide new genetic tools for metabolic engineering in Y. lipolytica and provide promoter engineering strategies that may be useful in engineering other non-model fungal systems.
['Shabbir Hussain|Murtaza|M|', 'Gambill|Lauren|L|', 'Smith|Spencer|S|', 'Blenner|Mark A|MA|']
D001483:Base Sequence D004742:Enhancer Elements, Genetic D005656:Fungal Proteins D005818:Genetic Engineering D010957:Plasmids D012319:RNA Polymerase II D060888:Real-Time Polymerase Chain Reaction D016385:TATA Box D025062:Yarrowia
promoter Yarrowia lipolytica enhancers POX2 UAS oleate response elements