Cks1 Is Required for G<sub>1</sub>Cyclin–Cyclin-Dependent Kinase Activity in Budding Yeast

Reynard, Gregory J.; Reynolds, William; Verma, Rati; Deshaies, Raymond J.

doi:10.1128/mcb.20.16.5858-5864.2000

Cited by 64 publications

(49 citation statements)

References 50 publications

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“…Cell division activity is controlled by the cell-cycle machinery where the activation of cyclin-dependent kinases by association with phase-specific cyclins controls the entry of the cell into different stages of the cell cycle (9). The poplar microarray contains EST homologues for a number of genes involved in cell-cycle control, like cyclins A, B, and H, CDC2, and CKS1 ( Fig.…”

Section: Resultsmentioning

confidence: 99%

A transcriptional roadmap to wood formation

Hertzberg

Aspeborg

Schrader

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

440

375

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The large vascular meristem of poplar trees with its highly organized secondary xylem enables the boundaries between different developmental zones to be easily distinguished. This property of wood-forming tissues allowed us to determine a unique tissuespecific transcript profile for a well defined developmental gradient. RNA was prepared from different developmental stages of xylogenesis for DNA microarray analysis by using a hybrid aspen unigene set consisting of 2,995 expressed sequence tags. The analysis revealed that the genes encoding lignin and cellulose biosynthetic enzymes, as well as a number of transcription factors and other potential regulators of xylogenesis, are under strict developmental stage-specific transcriptional regulation.T ranscript profiling has the potential to reveal transcriptional hierarchy during development for thousands of genes, as well as providing expression data for many genes of unknown function (1, 2). This is especially true when expression patterns can be obtained for well defined tissues at specific developmental stages. However, this is technically demanding and so far there are no reports demonstrating tissue-specific analysis on cell types from a single developmental sequence. We have studied the developing secondary xylem of poplar, which is highly organized with easily recognized and distinct boundaries between the different developmental stages. Wood formation is initiated in the vascular cambium. Cambial derivatives develop into xylem cells through the processes of division, expansion, secondary wall formation, lignification, and finally, programmed cell death. The large physical size of the vascular meristem in trees offers a unique possibility to obtain samples from defined developmental stages by tangential cryo sectioning (3). To determine the steady-state mRNA levels at specific stages during the ontogeny of wood formation in Populus tremula ϫ Populus tremuloides (hybrid aspen) we sampled 30-m-thick sections through the wood development region and subsequently analyzed the samples by using a spotted cDNA-microarray (4) consisting of 2,995 unique ESTs from hybrid aspen. Our study provides a unique global examination of gene expression patterns that encompasses a developmental gradient within a multicellular organism. Materials and MethodsThe Unigene set was selected from the expressed sequence tags (ESTs) presented in ref. 5, using cluster analysis. ESTs were transformed into Escherichia coli by using TSS competent cells (6), plasmids were prepared by using 96-well Multiscreen FB plates (Millipore), inserts were PCR amplified by using vectorspecific primers, and PCR products were purified on Multiscreen PCR filter plates (Millipore) and spotted in duplicate onto CMT GAPS slides (Corning) by using the GMS 417 Arrayer (Affymetrix, Santa Clara, CA) as described (7). All PCR products were checked on ethidium bromide-stained agarose gels. Nine clones giving double PCR bands were excluded from the analysis.A subset of 2,085 of the 2,995 PCR products in the Unigene set...

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Section: Resultsmentioning

confidence: 99%

A transcriptional roadmap to wood formation

Hertzberg

Aspeborg

Schrader

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

440

375

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“…Optimal kinase activity of Cdc28 has been shown to require Cks1, at least in conjunction with G 1 cyclins (39). To eliminate the possibility that cyclins other than Clb6 were unable to stimulate phosphorylation of Swi6 because of limiting Cks1 activity, kinase assays with Cdc28 and individual cyclins were repeated with additional affinity-purified Cks1 (Fig.…”

Section: Resultsmentioning

confidence: 99%

Clb6/Cdc28 and Cdc14 Regulate Phosphorylation Status and Cellular Localization of Swi6

Geymonat¹,

Spanos²,

Wells³

et al. 2004

Molecular and Cellular Biology

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Nuclear export of the transcription factor Swi6 during the budding yeast Saccharomyces cerevisiae cell cycle is known to require phosphorylation of the Swi6 serine 160 residue. We show that Clb6/Cdc28 kinase is required for this nuclear export. Furthermore, Cdc28 combined with the S-phase cyclin Clb6 specifically phosphorylates serine 160 of Swi6 in vitro. Nuclear import of Swi6 occurs concomitantly with dephosphorylation of serine 160 in late M phase. We show that Cdc14 phosphatase, the principal effector of the mitotic exit network, can trigger nuclear import of Swi6 in vivo and that Cdc14 dephosphorylates Swi6 at serine 160 in vitro. Taken together, these observations show how Swi6 dephosphorylation and phosphorylation are integrated into changes of Cdc28 activity governing entry and exit from the G 1 phase of the cell cycle.

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“…Cks1 was originally discovered as an essential gene product in yeast that forms a complex with Cdks (19,20). In addition to binding to Cdk and regulating substrate selection and extent of substrate phosphorylation (21)(22)(23), Cks1 has been shown to be required for ubiquitination of p27 (17,18). Cks1, but not its closely related protein Cks2, binds to Skp2 (18).…”

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A Negatively Charged Amino Acid in Skp2 Is Required for Skp2-Cks1 Interaction and Ubiquitination of p27Kip1

Wang¹,

Ungermannova²,

Chen

et al. 2003

Journal of Biological Chemistry

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Proteolysis of cyclin-dependent kinase inhibitor p27 occurs predominately in the late G 1 phase of the cell cycle through a ubiquitin-mediated protein degradation pathway. Ubiquitination of p27 requires the SCF Skp2 ubiquitin ligase and Skp2 F-box binding protein Cks1. The mechanisms by which Skp2 recognizes Cks1 to ubiquitylate p27 remain obscure. Here we show that Asp-331 in the carboxyl terminus of Skp2 is required for its association with Cks1 and ubiquitination of p27. Mutation of Asp-331 to Ala disrupts the interaction between Skp2 and Cks1. Although Asp-331 mutation negates the ability of the Skp1-Cullin-F-box protein (SCF) complex to ubiquitylate p27, such a mutation has no effect on Skp2 selfubiquitination. A conservative change from Asp to Glu at position 331 of Skp2 does not affect Skp2-Cks1 interaction. Our results revealed a unique requirement for a negatively charged residue in the carboxyl-terminal region of Skp2 in recognition of Cks1 and ubiquitination of p27.

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Cks1 Is Required for G₁Cyclin–Cyclin-Dependent Kinase Activity in Budding Yeast

Cited by 64 publications

References 50 publications

A transcriptional roadmap to wood formation

A transcriptional roadmap to wood formation

Clb6/Cdc28 and Cdc14 Regulate Phosphorylation Status and Cellular Localization of Swi6

A Negatively Charged Amino Acid in Skp2 Is Required for Skp2-Cks1 Interaction and Ubiquitination of p27Kip1

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Cks1 Is Required for G1Cyclin–Cyclin-Dependent Kinase Activity in Budding Yeast

Cited by 64 publications

References 50 publications

A transcriptional roadmap to wood formation

A transcriptional roadmap to wood formation

Clb6/Cdc28 and Cdc14 Regulate Phosphorylation Status and Cellular Localization of Swi6

A Negatively Charged Amino Acid in Skp2 Is Required for Skp2-Cks1 Interaction and Ubiquitination of p27Kip1

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Cks1 Is Required for G₁Cyclin–Cyclin-Dependent Kinase Activity in Budding Yeast