The NDR/LATS family kinase Cbk1 directly controls transcriptional asymmetry.

PLoS Biology
Emily MazankaEric L Weiss

Abstract

Cell fate can be determined by asymmetric segregation of gene expression regulators. In the budding yeast Saccharomyces cerevisiae, the transcription factor Ace2 accumulates specifically in the daughter cell nucleus, where it drives transcription of genes that are not expressed in the mother cell. The NDR/LATS family protein kinase Cbk1 is required for Ace2 segregation and function. Using peptide scanning arrays, we determined Cbk1's phosphorylation consensus motif, the first such unbiased approach for an enzyme of this family, showing that it is a basophilic kinase with an unusual preference for histidine -5 to the phosphorylation site. We found that Cbk1 phosphorylates such sites in Ace2, and that these modifications are critical for Ace2's partitioning and function. Using proteins marked with GFP variants, we found that Ace2 moves from isotropic distribution to the daughter cell nuclear localization, well before cytokinesis, and that the nucleus must enter the daughter cell for Ace2 accumulation to occur. We found that Cbk1, unlike Ace2, is restricted to the daughter cell. Using both in vivo and in vitro assays, we found that two critical Cbk1 phosphorylations block Ace2's interaction with nuclear export machinery, while a t...Continue Reading

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Citations

Feb 5, 2014·The Journal of Cell Biology·Jessica ZapataDouglas R Kellogg
Sep 19, 2008·PLoS Biology·Emily J Parnell, David J Stillman
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Nov 21, 2018·The Journal of Cell Biology·Jennifer L BraceEric L Weiss

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Methods Mentioned

BETA
immunoprecipitation
ChIP
fluorescence microscopy
pulldown
Fluorescence
PCR
peptide library array
pulldowns

Software Mentioned

Odyssey
Openlab

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