Phosphorylation of the yeast nitrate transporter Ynt1 is essential for delivery to the plasma membrane during nitrogen limitation.

The Journal of Biological Chemistry
Francisco J NavarroJosé M Siverio

Abstract

Ynt1 is the sole high affinity nitrate transporter of the yeast Hansenula polymorpha. It is highly regulated by the nitrogen source, by being down-regulated in response to glutamine by repression of the YNT1 gene and Ynt1 ubiquitinylation, endocytosis, and vacuolar degradation. On the contrary, we show that nitrogen limitation stabilizes Ynt1 levels at the plasma membrane, requiring phosphorylation of the transporter. We determined that Ser-246 in the central intracellular loop plays a key role in the phosphorylation of Ynt1 and that the nitrogen permease reactivator 1 kinase (Npr1) is necessary for Ynt1 phosphorylation. Abolition of phosphorylation led Ynt1 to the vacuole by a pep12-dependent end4-independent pathway, which is also dependent on ubiquitinylation, whereas Ynt1 protein lacking ubiquitinylation sites does not follow this pathway. We found that, under nitrogen limitation, Ynt1 phosphorylation is essential for rapid induction of nitrate assimilation genes. Our results suggest that, under nitrogen limitation, phosphorylation prevents Ynt1 delivery from the secretion route to the vacuole, which, aided by reduced ubiquitinylation, accumulates Ynt1 at the plasma membrane. This mechanism could be part of the response tha...Continue Reading

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Citations

Dec 7, 2011·Proceedings of the National Academy of Sciences of the United States of America·Juan A BonachelaSimon A Levin
Jun 10, 2010·The Journal of Biological Chemistry·Nakako Shibagaki, Arthur R Grossman
Aug 7, 2013·Journal of Experimental Botany·Emanuel Sanz-LuqueEmilio Fernandez
May 26, 2010·Plant Physiology·Wirulda PootakhamArthur R Grossman
Sep 22, 2009·Journal of Microbiological Methods·Weidong QianBingsheng Qiu
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Dec 24, 2013·Eukaryotic Cell·Elisa CabreraJosé M Siverio
Apr 4, 2017·Journal of Experimental Botany·Aurore JacquotLaurence Lejay

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