Cyclin-dependent kinase (CDK) phosphorylation destabilizes somatic Wee1 via multiple pathways

Watanabe, Nobumoto; Arai, Harumi; Iwasaki, Jun-ichi; Shiina, Masaaki; Ogata, Kazuhiro; Hunter, Tony; Osada, Hiroyuki

doi:10.1073/pnas.0500410102

Cited by 190 publications

(204 citation statements)

References 32 publications

Supporting

Mentioning

190

Contrasting

Unclassified

Order By: Relevance

“…PBDdependent binding was examined by GST pulldown assay as described (9). PPG was added to bacterial lysates before mixing with lysates of HeLa cells expressing Wee1A.…”

Section: Methodsmentioning

confidence: 99%

“…At this stage, Plk1 also regulates mitotic entry through the phosphorylation and activation of Cdc25 (6,7). In parallel, the phosphorylation of Wee1 by Plk1, which is primed by Cdk1, promotes the destruction of Wee1, the inhibitory kinase of mitotic entry (8,9). By metaphase, a fraction of Plk1 relocalizes to the kinetochores, where it is involved in the regulation of the spindle assembly checkpoint pathway and proper chromosome segregation.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Deficiency in Chromosome Congression by the Inhibition of Plk1 Polo Box Domain-dependent Recognition

Watanabe

Sekine

Takagi

et al. 2009

Journal of Biological Chemistry

Self Cite

105

View full text Add to dashboard Cite

Polo-like kinase 1 (Plk1) is one of the key regulators of mitotic cell division. In addition to an N-terminal protein kinase catalytic domain, Plk1 possesses a phosphopeptide binding domain named polo box domain (PBD) at its C terminus. PBD is postulated to be essential for Plk1 localization and substrate targeting. Here, we developed a high-throughput screening system to identify inhibitors of PBD-dependent binding and screened a chemical library. We isolated a benzotropolone-containing natural compound derived from nutgalls (purpurogallin (PPG)) that inhibited PBD-dependent binding in vitro and in vivo. PPG not only delayed the onset of mitosis but also prolonged the progression of mitosis in HeLa cells. Although apparently normal bipolar spindles were formed even in the presence of PPG, the perturbation of chromosome alignment at metaphase plates activated the spindle assembly checkpoint pathway. These results demonstrate the predominant role of PBD-dependent binding on smooth chromosome congression at metaphase. In eukaryotes, Plk12 (or its orthologs) possesses a wide variety of essential functions during mitosis (1, 2). Although levels of Plk1 increase in late G 2 and rapidly decrease by proteolysis at the end of M phase, Plk1 localization changes dramatically when cells transit through the various mitotic stages. During late G 2 and prophase, Plk1 localizes primarily at the centrosomes, where Plk1 is involved in centrosome maturation, separation, and microtubule nucleation (3-5). At this stage, Plk1 also regulates mitotic entry through the phosphorylation and activation of Cdc25 (6, 7). In parallel, the phosphorylation of Wee1 by Plk1, which is primed by Cdk1, promotes the destruction of Wee1, the inhibitory kinase of mitotic entry (8, 9). By metaphase, a fraction of Plk1 relocalizes to the kinetochores, where it is involved in the regulation of the spindle assembly checkpoint pathway and proper chromosome segregation. Plk1 sensitizes the tension between sister chromatids and regulates the stability of kinetochore-microtubule interactions by phosphorylating kinetochore proteins such as BubR1 and proteins that harbor 3F3/2 epitopes (4, 10 -14). Although Plk1 localization at chromosome arms is required for loss of arm cohesion during M-phase (15, 16), localized Plk1 activity at the kinetochore is important for the proper accumulation of kinetochore proteins such as BubR1, Mad1, Mad2, Cdc20, and centromere/ kinetochore-associated protein-E (CENP-E) (4, 10 -12, 17, 18). During anaphase, Plk1 is concentrated in the spindle midzone, where it may facilitate microtubule sliding. After chromosomal segregation, Plk1 remains in the central spindle and midbody, where it is involved in formation of the cleavage furrow (19 -21).These various localizations of Plk1 are mediated by its noncatalytic C-terminal half, the polo box domain (PBD), which comprises two polo box motifs (22, 23). Recently, Yaffe and co-workers (24, 25) discovered that the PBD constitutes a phosphopeptide binding domain which binds with maximal affin...

show abstract

“…PBDdependent binding was examined by GST pulldown assay as described (9). PPG was added to bacterial lysates before mixing with lysates of HeLa cells expressing Wee1A.…”

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Deficiency in Chromosome Congression by the Inhibition of Plk1 Polo Box Domain-dependent Recognition

Watanabe

Sekine

Takagi

et al. 2009

Journal of Biological Chemistry

Self Cite

105

View full text Add to dashboard Cite

show abstract

“…Phosphorylation of the N-terminal Wee1 residues serines 53 and 123 are important for its turnover (9,14). Given the locale of leucine 483 within the Wee1 kinase activation domain (residues 461-488) (20), we postulated that this domain was necessary for Wee1 destruction and that degradation might be mediated by phosphorylation of serines 471, 472, and/or 480 that reside within this domain.…”

Section: Identified Wee1 Mutations Inhibit Wee1 Turnover In Vitro-mentioning

confidence: 99%

“…Wee1 degradation has been observed in Saccharomyces cerevisiae, Xenopus egg extracts, and somatic cells (5)(6)(7)(8)(9)(10)(11). In Xenopus, the nuclei are required for proper Wee1 degradation, and the completion of DNA replication is required to achieve maximal rates of Wee1 degradation (6).…”

mentioning

confidence: 99%

Activation Domain-dependent Degradation of Somatic Wee1 Kinase

Owens

Simanski

Squire

et al. 2010

Journal of Biological Chemistry

View full text Add to dashboard Cite

Cell cycle progression is dependent upon coordinate regulation of kinase and proteolytic pathways. Inhibitors of cell cycle transitions are degraded to allow progression into the subsequent cell cycle phase. For example, the tyrosine kinase and Cdk1 inhibitor Wee1 is degraded during G 2 and mitosis to allow mitotic progression. Previous studies suggested that the N terminus of Wee1 directs Wee1 destruction. Using a chemical mutagenesis strategy, we report that multiple regions of Wee1 control its destruction. Most notably, we find that the activation domain of the Wee1 kinase is also required for its degradation. Mutations in this domain inhibit Wee1 degradation in somatic cell extracts and in cells without affecting the overall Wee1 structure or kinase activity. More broadly, these findings suggest that kinase activation domains may be previously unappreciated sites of recognition by the ubiquitin proteasome pathway.The tyrosine kinase Wee1 inhibits mitotic entry, and its degradation is essential for exit from the G 2 phase of the cell cycle (1). Here, Wee1 inactivates the mitosis-specific cyclin-dependent kinase Cdk1-cyclin B complex during the S and G 2 phases by phosphorylating Cdk1 at tyrosine 15. However, Wee1 activity is opposed by the phosphatase Cdc25, which removes phosphate from tyrosine 15 on Cdk1, thereby activating Cdk1-cyclin B at the G 2 -M transition (2). A major mechanism that tips the balance toward Cdc25 and mitotic entry is Wee1 degradation during G 2 , and as this occurs active Cdc25 and Cdk1-cyclin B form a positive feedback loop, which ensures that mitotic entry is unidirectional (3, 4). Thus, Wee1 degradation is an essential component of the Cdk1 activation circuit.Wee1 degradation has been observed in Saccharomyces cerevisiae, Xenopus egg extracts, and somatic cells (5-11). In Xenopus, the nuclei are required for proper Wee1 degradation, and the completion of DNA replication is required to achieve maximal rates of Wee1 degradation (6). Thus, Wee1 degradation is part of a sensing mechanism that signals the completion of the DNA replication phase, ensuring proper timing of entry into mitosis. For example, when DNA replication stalls, this fail-safe mechanism allows defects to be corrected before cells enter mitosis. Indeed, current theories suggest that many cancer cells have ineffective checkpoint pathways that cause them to divide with incompletely replicated DNA, which leads to genomic instability (12). Consistent with this notion are the findings that wee1 knock-out mice are not viable and that wee1 Ϫ/Ϫ cells undergo mitotic catastrophe (13).Wee1 phosphorylation is required for its degradation and is regulated by the DNA replication checkpoint (6, 10). Here in the presence of the DNA replication checkpoint, Wee1 phosphorylation is negligible, whereas when DNA replication proceeds normally, Wee1 phosphorylation and degradation and mitotic entry are normal.One aspect of Wee1 degradation conserved between embryonic and somatic cell cycles is the requirement for SCF 3 ubiquitin ligases th...

show abstract

“…Thus, entry into M phase requires both inhibition of Wee1-related kinases and activation of Cdc25 phosphatases (figure 2a). This is brought about by Polo-like kinase 1 (Plk1; Plo1 in fission yeast), which phosphorylates both Cdc25 and Wee1 [37][38][39][40], with the result that Cdc25 is activated whilst Wee1 is inhibited-in fact targeted for degradation [41] ( figure 2a,c). Importantly, the recruitment of Plk1 to both Wee1 and Cdc25 depends on prior phosphorylation of these enzymes by an initial pool of activated Cdk1.…”

Section: Introductionmentioning

confidence: 99%

Centrosomes as signalling centres

Arquint

Gabryjonczyk

Nigg

2014

Phil. Trans. R. Soc. B

126

114

View full text Add to dashboard Cite

Centrosomes—as well as the related spindle pole bodies (SPBs) of yeast—have been extensively studied from the perspective of their microtubule-organizing roles. Moreover, the biogenesis and duplication of these organelles have been the subject of much attention, and the importance of centrosomes and the centriole–ciliary apparatus for human disease is well recognized. Much less developed is our understanding of another facet of centrosomes and SPBs, namely their possible role as signalling centres. Yet, many signalling components, including kinases and phosphatases, have been associated with centrosomes and spindle poles, giving rise to the hypothesis that these organelles might serve as hubs for the integration and coordination of signalling pathways. In this review, we discuss a number of selected studies that bear on this notion. We cover different processes (cell cycle control, development, DNA damage response) and organisms (yeast, invertebrates and vertebrates), but have made no attempt to be comprehensive. This field is still young and although the concept of centrosomes and SPBs as signalling centres is attractive, it remains primarily a concept—in need of further scrutiny. We hope that this review will stimulate thought and experimentation.

show abstract

Cyclin-dependent kinase (CDK) phosphorylation destabilizes somatic Wee1 via multiple pathways

Cited by 190 publications

References 32 publications

Deficiency in Chromosome Congression by the Inhibition of Plk1 Polo Box Domain-dependent Recognition

Deficiency in Chromosome Congression by the Inhibition of Plk1 Polo Box Domain-dependent Recognition

Activation Domain-dependent Degradation of Somatic Wee1 Kinase

Centrosomes as signalling centres

Contact Info

Product

Resources

About