Oligomers of the Cdc7/Dbf4 protein kinase exist in the yeast cell

Shellman, Yiqun G.; Schauer, Irene E.; Oshiro, Guy; Dohrmann, Paul R.; Sclafani, Robert A.

doi:10.1007/s004380050833

Cited by 20 publications

(15 citation statements)

References 20 publications

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“…In this case, the ability of two different alleles to form dimers may be the determinant of complementation. Interallelic complementation based on dimerization has been observed with Cdc7p (Shellman et al, 1998). However, our two-hybrid and coimmunoprecipitation tests were unable to detect interaction between Cdc28p monomers.…”

Section: Multiple Functions Of Cdc28p Affecting Mitotic Progression Acontrasting

confidence: 55%

“…Although heteroallelic combinations of Ala55Thr and Ile59Asn yielded highly polarized cells (Figure 4), indicating noncomplementation, other pairs of alleles displayed complementation for the enhanced cell polarity and G2/M-shift phenotypes (Figure 4; Table 2). This pattern of complementation suggested that Cdc28p might function as a dimer as had been observed for Cdc7p (Shellman et al, 1998). To test this possibility, the CDC28 ORF was cloned into bait-and-prey yeast two-hybrid expression vectors in each of two experimental systems.…”

Section: The Cdc28 Ecp Mutants Define Multiple Complementation Groupsmentioning

confidence: 87%

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Enhanced Cell Polarity in Mutants of the Budding Yeast Cyclin-dependent Kinase Cdc28p

Ahn

Tobe²,

Gerald

et al. 2001

MBoC

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The yeast cyclin-dependent kinase Cdc28p regulates bud morphogenesis and cell cycle progression via the antagonistic activities of Cln and Clb cyclins. Cln G1 cyclins direct polarized growth and bud emergence, whereas Clb G2 cyclins promote isotropic growth of the bud and chromosome segregation. Using colony morphology as a screen to dissect regulation of polarity by Cdc28p, we identified nine point mutations that block the apical-isotropic switch while maintaining other functions. Like a clb2⌬ mutation, each confers tubular bud shape, apically polarized actin distribution, unipolar budding, and delayed anaphase. The mutations are all suppressed by CLB2 overexpression and are synthetically lethal with a CLB2 deletion. However, defects in multiple independent pathways may underlie their common phenotype, because the mutations are scattered throughout the CDC28 sequence, complement each other, and confer diverse biochemical properties. Glu12Gly, a mutation that alters a residue involved in Swe1p inhibition of Cdc28p, was unique in being suppressed by deficiency of SWE1 or CLN1. With wild-type CDC28, filament formation induced by CLN1 overexpression was markedly decreased in a SWE1 deletion. These results suggest that Swe1p, via inhibition of Clb2p/Cdc28p, may mediate much of the effect of Cln1p on filamentous morphogenesis.

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Section: Multiple Functions Of Cdc28p Affecting Mitotic Progression Acontrasting

confidence: 55%

Section: The Cdc28 Ecp Mutants Define Multiple Complementation Groupsmentioning

confidence: 87%

Enhanced Cell Polarity in Mutants of the Budding Yeast Cyclin-dependent Kinase Cdc28p

Ahn

Tobe²,

Gerald

et al. 2001

MBoC

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“…DDK or Dbf4 oligomerization has been suggested previously (Shellman et al 1998;Matthews et al 2012). Furthermore, tandem BRCT domains form dimers that bind phospho-S/T motifs (Caldecott 2003;Rodriguez and Songyang 2008), and intermolecular dimerization between BRCT domains has also been described for the DNA repair proteins XRCC1 and Ligase III (Cuneo et al 2011).…”

Section: Models For Rad53 Binding To Dbf4mentioning

confidence: 88%

DNA Replication Checkpoint Signaling Depends on a Rad53–Dbf4 N-Terminal Interaction in Saccharomyces cerevisiae

et al. 2013

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Dbf4-dependent kinase (DDK) and cyclin-dependent kinase (CDK) are essential to initiate DNA replication at individual origins. During replication stress, the S-phase checkpoint inhibits the DDK-and CDK-dependent activation of late replication origins. Rad53 kinase is a central effector of the replication checkpoint and both binds to and phosphorylates Dbf4 to prevent late-origin firing. The molecular basis for the Rad53-Dbf4 physical interaction is not clear but occurs through the Dbf4 N terminus. Here we found that both Rad53 FHA1 and FHA2 domains, which specifically recognize phospho-threonine (pT), interacted with Dbf4 through an N-terminal sequence and an adjacent BRCT domain. Purified Rad53 FHA1 domain (but not FHA2) bound to a pT Dbf4 peptide in vitro, suggesting a possible phospho-threonine-dependent interaction between FHA1 and Dbf4. The Dbf4-Rad53 interaction is governed by multiple contacts that are separable from the Cdc5-and Msa1-binding sites in the Dbf4 N terminus. Importantly, abrogation of the Rad53-Dbf4 physical interaction blocked Dbf4 phosphorylation and allowed late-origin firing during replication checkpoint activation. This indicated that Rad53 must stably bind to Dbf4 to regulate its activity.T HE fidelity of chromosome replication depends on checkpoint mechanisms to stabilize stalled forks, regulate origin activation, and repair DNA damage (Hartwell and Weinert 1989;Bartek et al. 2004;Segurado and Tercero 2009). In response to replication stress, the replication checkpoint maintains replisome stability and prevents late origins from firing, which allows time for DNA repair and the completion of DNA replication prior to chromosome segregation. Incomplete DNA replication or uncoordinated origin firing following DNA damage can result in genomic instability, cancer predisposition, and premature aging (Branzei and Foiani 2010).In the budding yeast Saccharomyces cerevisiae, activation of the checkpoint sensor kinase Mec1 (vertebrate ATR, Ataxia Telangiectasia and Rad3-related) is triggered at stalled forks or sites of DNA damage (Majka et al. 2006;Labib and De Piccoli 2011). Subsequent signal amplification through the Mrc1 or Rad9 adaptors leads to activation of the checkpoint kinase Rad53 (the ortholog of the human tumor suppressor Chk2) (Branzei and Foiani 2009). Rad53 is an integral transducer of various cellular responses to replication stress or DNA damage. Rad53 induces a series of transcriptional responses through MBF-regulated genes (Bastos de Oliveira et al. 2012;Travesa et al. 2012) and also activates the Dun1 kinase, which promotes the expression of ribonucleotide reductase (RNR) subunits and additional DNA repair genes (Huang et al. 1998). In parallel, Rad53 down-regulates the RNR inhibitor Sml1 to increase deoxyribonucleotide levels and facilitate DNA synthesis (Zhao et al. 2001). In response to replication fork stalling, Rad53 prevents the activation of late replication origins by phosphorylating two proteins required for the initiation of DNA replication: Dbf4 and Sld3 (...

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“…One-hybrid and two-hybrid assays: Liquid culture b-galactosidase and X-Gal assays were done as previously described (Shellman et al 1998;Dohrmann et al 1999).…”

Section: Methodsmentioning

confidence: 99%

Novel Role for Checkpoint Rad53 Protein Kinase in the Initiation of Chromosomal DNA Replication inSaccharomyces cerevisiae

Dohrmann

Sclafani

2006

Genetics

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A novel role for Rad53 in the initiation of DNA replication that is independent of checkpoint or deoxynucleotide regulation is proposed. Rad53 kinase is part of a signal transduction pathway involved in the DNA damage and replication checkpoints, while Cdc7-Dbf4 kinase (DDK) is important for the initiation of DNA replication. In addition to the known cdc7-rad53 synthetic lethality, rad53 mutations suppress mcm5-bob1, a mutation in the replicative MCM helicase that bypasses DDK's essential role. Rad53 kinase activity but neither checkpoint FHA domain is required. Conversely, Rad53 kinase can be activated without DDK. Rad53's role in replication is independent of both DNA and mitotic checkpoints because mutations in other checkpoint genes that act upstream or downstream of RAD53 or in the mitotic checkpoint do not exhibit these phenotypes. Because Rad53 binds an origin of replication mainly through its kinase domain and rad53 null mutants display a minichromosome loss phenotype, Rad53 is important in the initiation of DNA replication, as are DDK and Mcm2-7 proteins. This unique requirement for Rad53 can be suppressed by the deletion of the major histone H3/H4 gene pair, indicating that Rad53 may be regulating initiation by controlling histone protein levels and/or by affecting origin chromatin structure.

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Oligomers of the Cdc7/Dbf4 protein kinase exist in the yeast cell

Cited by 20 publications

References 20 publications

Enhanced Cell Polarity in Mutants of the Budding Yeast Cyclin-dependent Kinase Cdc28p

Enhanced Cell Polarity in Mutants of the Budding Yeast Cyclin-dependent Kinase Cdc28p

DNA Replication Checkpoint Signaling Depends on a Rad53–Dbf4 N-Terminal Interaction in Saccharomyces cerevisiae

Novel Role for Checkpoint Rad53 Protein Kinase in the Initiation of Chromosomal DNA Replication inSaccharomyces cerevisiae

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