The NDR/LATS family kinase Cbk1 directly controls transcriptional asymmetry.
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. T...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2008-08-01
|
| Series: | PLoS Biology |
| Online Access: | http://europepmc.org/articles/PMC2517623?pdf=render |
| _version_ | 1818905534886051840 |
|---|---|
| author | Emily Mazanka Jess Alexander Brian J Yeh Patrick Charoenpong Drew M Lowery Michael Yaffe Eric L Weiss |
| author_facet | Emily Mazanka Jess Alexander Brian J Yeh Patrick Charoenpong Drew M Lowery Michael Yaffe Eric L Weiss |
| author_sort | Emily Mazanka |
| collection | DOAJ |
| description | 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 third distal modification most likely acts to increase the transcription factor's activity. Our findings show that Cbk1 directly controls Ace2, regulating the transcription factor's activity and interaction with nuclear export machinery through three phosphorylation sites. Furthermore, Cbk1 exhibits a novel specificity that is likely conserved among related kinases from yeast to metazoans. Cbk1 is functionally restricted to the daughter cell, and cannot diffuse from the daughter to the mother. In addition to providing a mechanism for Ace2 segregation, these findings show that an isotropically distributed cell fate determinant can be asymmetrically partitioned in cytoplasmically contiguous cells through spatial segregation of a regulating protein kinase. |
| first_indexed | 2024-12-19T21:24:52Z |
| format | Article |
| id | doaj.art-b6f0d5532677435fbdc486a3e45e5dcf |
| institution | Directory Open Access Journal |
| issn | 1544-9173 1545-7885 |
| language | English |
| last_indexed | 2024-12-19T21:24:52Z |
| publishDate | 2008-08-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Biology |
| spelling | doaj.art-b6f0d5532677435fbdc486a3e45e5dcf2022-12-21T20:05:10ZengPublic Library of Science (PLoS)PLoS Biology1544-91731545-78852008-08-0168e20310.1371/journal.pbio.0060203The NDR/LATS family kinase Cbk1 directly controls transcriptional asymmetry.Emily MazankaJess AlexanderBrian J YehPatrick CharoenpongDrew M LoweryMichael YaffeEric L WeissCell 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 third distal modification most likely acts to increase the transcription factor's activity. Our findings show that Cbk1 directly controls Ace2, regulating the transcription factor's activity and interaction with nuclear export machinery through three phosphorylation sites. Furthermore, Cbk1 exhibits a novel specificity that is likely conserved among related kinases from yeast to metazoans. Cbk1 is functionally restricted to the daughter cell, and cannot diffuse from the daughter to the mother. In addition to providing a mechanism for Ace2 segregation, these findings show that an isotropically distributed cell fate determinant can be asymmetrically partitioned in cytoplasmically contiguous cells through spatial segregation of a regulating protein kinase.http://europepmc.org/articles/PMC2517623?pdf=render |
| spellingShingle | Emily Mazanka Jess Alexander Brian J Yeh Patrick Charoenpong Drew M Lowery Michael Yaffe Eric L Weiss The NDR/LATS family kinase Cbk1 directly controls transcriptional asymmetry. PLoS Biology |
| title | The NDR/LATS family kinase Cbk1 directly controls transcriptional asymmetry. |
| title_full | The NDR/LATS family kinase Cbk1 directly controls transcriptional asymmetry. |
| title_fullStr | The NDR/LATS family kinase Cbk1 directly controls transcriptional asymmetry. |
| title_full_unstemmed | The NDR/LATS family kinase Cbk1 directly controls transcriptional asymmetry. |
| title_short | The NDR/LATS family kinase Cbk1 directly controls transcriptional asymmetry. |
| title_sort | ndr lats family kinase cbk1 directly controls transcriptional asymmetry |
| url | http://europepmc.org/articles/PMC2517623?pdf=render |
| work_keys_str_mv | AT emilymazanka thendrlatsfamilykinasecbk1directlycontrolstranscriptionalasymmetry AT jessalexander thendrlatsfamilykinasecbk1directlycontrolstranscriptionalasymmetry AT brianjyeh thendrlatsfamilykinasecbk1directlycontrolstranscriptionalasymmetry AT patrickcharoenpong thendrlatsfamilykinasecbk1directlycontrolstranscriptionalasymmetry AT drewmlowery thendrlatsfamilykinasecbk1directlycontrolstranscriptionalasymmetry AT michaelyaffe thendrlatsfamilykinasecbk1directlycontrolstranscriptionalasymmetry AT ericlweiss thendrlatsfamilykinasecbk1directlycontrolstranscriptionalasymmetry AT emilymazanka ndrlatsfamilykinasecbk1directlycontrolstranscriptionalasymmetry AT jessalexander ndrlatsfamilykinasecbk1directlycontrolstranscriptionalasymmetry AT brianjyeh ndrlatsfamilykinasecbk1directlycontrolstranscriptionalasymmetry AT patrickcharoenpong ndrlatsfamilykinasecbk1directlycontrolstranscriptionalasymmetry AT drewmlowery ndrlatsfamilykinasecbk1directlycontrolstranscriptionalasymmetry AT michaelyaffe ndrlatsfamilykinasecbk1directlycontrolstranscriptionalasymmetry AT ericlweiss ndrlatsfamilykinasecbk1directlycontrolstranscriptionalasymmetry |