Interaction of Yna1 and Yna2 Is Required for Nuclear Accumulation and Transcriptional Activation of the Nitrate Assimilation Pathway in the Yeast Hansenula polymorpha

PloS One
Lucia SilvestriniJoseph Strauss

Abstract

A few yeasts, including Hansenula polymorpha are able to assimilate nitrate and use it as nitrogen source. The genes necessary for nitrate assimilation are organised in this organism as a cluster comprising those encoding nitrate reductase (YNR1), nitrite reductase (YNI1), a high affinity transporter (YNT1), as well as the two pathway specific Zn(II)2Cys2 transcriptional activators (YNA1, YNA2). Yna1p and Yna2p mediate induction of the system and here we show that their functions are interdependent. Yna1p activates YNA2 as well as its own (YNA1) transcription thus forming a nitrate-dependent autoactivation loop. Using a split-YFP approach we demonstrate here that Yna1p and Yna2p form a heterodimer independently of the inducer and despite both Yna1p and Yna2p can occupy the target promoter as mono- or homodimer individually, these proteins are transcriptionally incompetent. Subsequently, the transcription factors target genes containing a conserved DNA motif (termed nitrate-UAS) determined in this work by in vitro and in vivo protein-DNA interaction studies. These events lead to a rearrangement of the chromatin landscape on the target promoters and are associated with the onset of transcription of these target genes. In contrast...Continue Reading

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Citations

Feb 26, 2019·Molecular Biotechnology·Roghayyeh BaghbanMaryam Aria
Sep 6, 2018·Annual Review of Genetics·Hans-Wilhelm NützmannAnne Osbourn

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Datasets Mentioned

BETA
AJ223294
AJ011906

Methods Mentioned

BETA
ubiquitination
histone acetylation
PCR
footprinting
confocal microscopy
immunoprecipitation
ChIP
electric mobility shift
MNase
fluorescence microscopy

Software Mentioned

Molecular Analyst / Macintosh Software
ImageQuant

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