Abstract
Greatwall (Gwl) functions as an essential mitotic kinase by antagonizing protein phosphatase 2A. In this study we identifi ed Hsp90, Cdc37 and members of the importin a and b families as the major binding partners of Gwl. Both Hsp90/Cdc37 chaperone and importin complexes associated with the N-terminal kinase domain of Gwl, whereas an intact glycine-rich loop at the N-terminus of Gwl was essential for binding of Hsp90/Cdc37 but not importins. We found that Hsp90 inhibition led to destabilization of Gwl, a mechanism that may partially contribute to the emerging role of Hsp90 in cell cycle progression and the anti-proliferative potential of Hsp90 inhibition. Moreover, in agreement with its importin association, Gwl exhibited nuclear localization in interphase Xenopus S3 cells, and dynamic nucleocytoplasmic distribution during mitosis. We identified KR456/457 as the locus of importin binding and the functional NLS of Gwl. Mutation of this site resulted in exclusion of Gwl from the nucleus. Finally, we showed that the Gwl nuclear localization is indispensable for the biochemical function of Gwl in promoting mitotic entry.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 3565-3575 |
| Number of pages | 11 |
| Journal | Cell Cycle |
| Volume | 13 |
| Issue number | 22 |
| DOIs | |
| State | Published - Nov 15 2014 |
Fingerprint
Keywords
- Greatwall kinase
- Hsp90
- Importin
- Mitosis
- Nuclear localization
ASJC Scopus subject areas
- Molecular Biology
- Developmental Biology
- Cell Biology
Cite this
Regulation of greatwall kinase by protein stabilization and nuclear localization. / Yamamoto, Tomomi M.; Wang, Ling; Fisher, Laura A.; Eckerdt, Frank D.; Peng, Aimin.
In: Cell Cycle, Vol. 13, No. 22, 15.11.2014, p. 3565-3575.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Regulation of greatwall kinase by protein stabilization and nuclear localization
AU - Yamamoto, Tomomi M.
AU - Wang, Ling
AU - Fisher, Laura A.
AU - Eckerdt, Frank D.
AU - Peng, Aimin
PY - 2014/11/15
Y1 - 2014/11/15
N2 - Greatwall (Gwl) functions as an essential mitotic kinase by antagonizing protein phosphatase 2A. In this study we identifi ed Hsp90, Cdc37 and members of the importin a and b families as the major binding partners of Gwl. Both Hsp90/Cdc37 chaperone and importin complexes associated with the N-terminal kinase domain of Gwl, whereas an intact glycine-rich loop at the N-terminus of Gwl was essential for binding of Hsp90/Cdc37 but not importins. We found that Hsp90 inhibition led to destabilization of Gwl, a mechanism that may partially contribute to the emerging role of Hsp90 in cell cycle progression and the anti-proliferative potential of Hsp90 inhibition. Moreover, in agreement with its importin association, Gwl exhibited nuclear localization in interphase Xenopus S3 cells, and dynamic nucleocytoplasmic distribution during mitosis. We identified KR456/457 as the locus of importin binding and the functional NLS of Gwl. Mutation of this site resulted in exclusion of Gwl from the nucleus. Finally, we showed that the Gwl nuclear localization is indispensable for the biochemical function of Gwl in promoting mitotic entry.
AB - Greatwall (Gwl) functions as an essential mitotic kinase by antagonizing protein phosphatase 2A. In this study we identifi ed Hsp90, Cdc37 and members of the importin a and b families as the major binding partners of Gwl. Both Hsp90/Cdc37 chaperone and importin complexes associated with the N-terminal kinase domain of Gwl, whereas an intact glycine-rich loop at the N-terminus of Gwl was essential for binding of Hsp90/Cdc37 but not importins. We found that Hsp90 inhibition led to destabilization of Gwl, a mechanism that may partially contribute to the emerging role of Hsp90 in cell cycle progression and the anti-proliferative potential of Hsp90 inhibition. Moreover, in agreement with its importin association, Gwl exhibited nuclear localization in interphase Xenopus S3 cells, and dynamic nucleocytoplasmic distribution during mitosis. We identified KR456/457 as the locus of importin binding and the functional NLS of Gwl. Mutation of this site resulted in exclusion of Gwl from the nucleus. Finally, we showed that the Gwl nuclear localization is indispensable for the biochemical function of Gwl in promoting mitotic entry.
KW - Greatwall kinase
KW - Hsp90
KW - Importin
KW - Mitosis
KW - Nuclear localization
UR - http://www.scopus.com/inward/record.url?scp=84919936035&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84919936035&partnerID=8YFLogxK
U2 - 10.4161/15384101.2014.962942
DO - 10.4161/15384101.2014.962942
M3 - Article
C2 - 25483093
AN - SCOPUS:84919936035
VL - 13
SP - 3565
EP - 3575
JO - Cell Cycle
JF - Cell Cycle
SN - 1538-4101
IS - 22
ER -