Requirement of Cdk2-Cyclin E Activity for Repeated Centrosome Reproduction in
            <i>Xenopus</i>
            Egg Extracts

Hinchcliffe, Edward H.; Li, Chuan; Thompson, E. A.; Maller, James L.; Sluder, Greenfield

doi:10.1126/science.283.5403.851

Cited by 481 publications

(389 citation statements)

References 32 publications

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“…Centrosome duplication and centriole separation require cyclin-dependent kinase (Cdk2) and can be speci®cally driven by Cdk2-cyclin E complexes. In the same way, p21 Waf-1 overexpression blocks both the duplication of centrosomes and centriole separation in di erent models (Hinchcli e et al, 1999;Lacey et al, 1999;Matsumoto et al, 1999). In SIAH-1 transfected cells, centrosome anomalies in number (in multinucleated cells), in migration and in morphology were found.…”

Section: Discussionmentioning

confidence: 73%

SIAH-1 inhibits cell growth by altering the mitotic process

et al. 1999

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SIAH-1, the human homologue of the drosophila seven in absentia gene, is a p53-p21 Waf-1 inducible gene. We report that stable transfection with SIAH-1 of the epithelial breast cancer cell line MCF-7 blocks its growth process. The transfectants show a redistribution of SIAH-1 protein within the nucleus, more speci®cally to the nuclear matrix, associated to dramatic changes in cell morphology and defective mitosis. Multinucleated giant cells (2 ± 12 nuclei in more than 50% cells) were a most striking observation associated with tubulin spindle disorganization and defective cytokinesis. There were also present at high frequency abortive mitotic ®gures, DNA bridges and persistance of intercellular bridges and midbodies, along with an increased expression of p21 Waf-1 . These results indicate that the mechanism of growth arrest induced by SIAH-1 in MCF-7 cells involves disorganization of the mitotic program, mainly during nuclei separation and cytokinesis.

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Section: Discussionmentioning

confidence: 73%

SIAH-1 inhibits cell growth by altering the mitotic process

et al. 1999

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“…Although cyclin A2 may not be the only transcriptional target relevant for this process, our results emphasize that ongoing cyclin A2 transcription is critical for centriole overduplication under conditions of an S phase arrest. The fact that cyclin A expression is predominantly affected by aamanitin in contrast to cyclin E may be related to the time of the HU-induced cell cycle arrest in S phase when cyclin A mRNA is synthesized and does not preclude that cyclin E plays a role in centriole overduplication (Hinchcliffe et al, 1999;Mussman et al, 2000). It is likely that cyclin A promotes centriole overduplication in S phase-arrested cells through a continued stimulation of CDK2 activity (Duensing et al, 2006).…”

Section: Discussionmentioning

confidence: 99%

RNA polymerase II transcription is required for human papillomavirus type 16 E7- and hydroxyurea-induced centriole overduplication

et al. 2006

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Aberrant centrosome numbers are detected in virtually all human cancers where they can contribute to chromosomal instability by promoting mitotic spindle abnormalities. Despite their widespread occurrence, the molecular mechanisms that underlie centrosome amplification are only beginning to emerge. Here, we present evidence for a novel regulatory circuit involved in centrosome overduplication that centers on RNA polymerase II (pol II). We found that human papillomavirus type 16 E7 (HPV-16 E7)-and hydroxyurea (HU)-induced centriole overduplication are abrogated by a-amanitin, a potent and specific RNA pol II inhibitor. In contrast, normal centriole duplication proceeded undisturbed in a-amanitin-treated cells. Centriole overduplication was significantly reduced by siRNA-mediated knock down of CREB-binding protein (CBP), a transcriptional co-activator. We identified cyclin A2 as a key transcriptional target of RNA pol II during HU-induced centriole overduplication. Collectively, our results show that ongoing RNA pol II transcription is required for centriole overduplication whereas it may be dispensable for normal centriole duplication. Given that many chemotherapeutic agents function through inhibition of transcription, our results may help to develop strategies to target centrosomemediated chromosomal instability for cancer therapy and prevention.

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“…Five reports using such methods have defined multiple roles of Cdk2/cyclin E in egg extracts. Cdk2/cyclin E is capable of providing all the Cdk activity necessary to support a single round of chromosomal DNA replication (Jackson et al, 1995;Strausfeld et al, 1996), plays a role in centrosome duplication (Hinchcliffe et al, 1999;Lacey et al, 1999), and is important for maintaining conditions permissive for cyclin B to activate Cdk1 and bring about mitosis (Guadagno and Newport, 1996). There are limits to the abilities of Cdk2/cyclin E, however, for it is unable to trigger entry into mitosis in the absence of other Cdk/cyclin complexes (Strausfeld et al, 1996), although a recent article claimed that it influenced the duration of M phase (D'Angiolella et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

Identification of the Nuclear Localization Signal inXenopusCyclin E and Analysis of Its Role in Replication and Mitosis

Moore

Kornbluth

Hunt

2002

MBoC

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Cyclin-dependent kinase (Cdk)2/cyclin E is imported into nuclei assembled in Xenopus egg extracts by a pathway that requires importin-␣ and -␤. Here, we identify a basic nuclear localization sequence (NLS) in the N-terminus of Xenopus cyclin E. Mutation of the NLS eliminated nuclear accumulation of both cyclin E and Cdk2, and such versions of cyclin E were unable to trigger DNA replication. Addition of a heterologous NLS from SV40 large T antigen restored both nuclear targeting of Cdk2/cyclin E and DNA replication. We present evidence indicating that Cdk2/cyclin E complexes must become highly concentrated within nuclei to support replication and find that cyclin A can trigger replication at much lower intranuclear concentrations. We confirmed that depletion of endogenous cyclin E increases the concentration of cyclin B necessary to promote entry into mitosis. In contrast to its inability to promote DNA replication, cyclin E lacking its NLS was able to cooperate with cyclin B in promoting mitotic entry. INTRODUCTIONCyclin-dependent kinases (Cdks) are vital for the initiation of both the major events of the eukaryotic cell cycle: the duplication of the genome in S phase and its segregation to two daughter cells during mitosis. In animal cells, several families of Cdks and cyclins have roles in cell cycle control. d-type cyclins complexed to Cdk4/Cdk6 regulate the decision to divide or differentiate, Cdk2/cyclin E and Cdk2/ cyclin A collaborate to initiate the events of S phase, and Cdk/cyclin A and Cdk1/cyclin B combine forces to trigger the wholesale reorganization of cellular components at mitosis (Girard et al., 1991;Pagano et al., 1992;Ohtsubo et al., 1995;Furuno et al., 1999;Hu et al., 2001;Nigg, 2001).Cdk/cyclin complexes are regulated by multiple mechanisms that ensure that they execute their functions at the correct time (Morgan, 1997); ubiquitylation and proteolysis of cyclins A and B are critical for M-phase exit (Wheatley et al., 1997;den Elzen and Pines, 2001; Geley et al., 2001), whereas defects in cyclin E proteolysis may be associated with certain cancers (Strohmaier et al., 2001). The subcellular localization of Cdk/cyclin complexes is also critical for faithful cell cycle control (Pines and Hunter, 1991;Hagting et al., 1998;Toyoshima et al., 1998;Pines, 1999;Alt et al., 2000;Draviam et al., 2001), and some progress has been made in identifying the mechanisms responsible, in particular those used by Cdk/cyclin complexes to shuttle between the cytoplasm and nucleus (Yang et al., 1998;Hagting et al., 1999;Moore et al., 1999;Takizawa et al., 1999). In this article, we focus attention on the mechanism and relevance of nuclear localization for the function of Cdk2/cyclin E in Xenopus egg extracts.Xenopus egg extracts have proved useful for studying the functions of Cdk/cyclin complexes in cell cycle control. Extracts exhibit excellent synchrony and faithfully recapitulate both S-phase and M-phase processes in vitro. Moreover, it is possible to manipulate their contents by depletion or addition of protei...

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Requirement of Cdk2-Cyclin E Activity for Repeated Centrosome Reproduction in Xenopus Egg Extracts

Cited by 481 publications

References 32 publications

SIAH-1 inhibits cell growth by altering the mitotic process

SIAH-1 inhibits cell growth by altering the mitotic process

RNA polymerase II transcription is required for human papillomavirus type 16 E7- and hydroxyurea-induced centriole overduplication

Identification of the Nuclear Localization Signal inXenopusCyclin E and Analysis of Its Role in Replication and Mitosis

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