cdt1 is an essential target of the Cdc10/Sct1 transcription factor: requirement for DNA replication and inhibition of mitosis.

Hofmann, Johannes; Beach, David

doi:10.1002/j.1460-2075.1994.tb06277.x

Cited by 223 publications

(196 citation statements)

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“…3, B and C, lanes and bars 13), double and triple mutations in these residues caused the reduced affinities toward Mcm4/6/7 proteins (Fig. 3, B and C, lanes and bars [15][16][17], which is qualitatively in agreement with the results of yeast viability assay. You and Masai (30) …”

Section: Regions Of Winged-helix Domain For Binding To MCMsupporting

confidence: 86%

“…Cdt1 was originally found in fission yeast (16), and its function as a factor involved in replication licensing was first characterized in Xenopus and fission yeast (17,18 Mcm2-7 both in vitro and in vivo (19 -22). Cdt1 can be divided into three functional regions, namely, N-terminal, central, and C-terminal regions, which have DNA-, geminin-, and Mcm2-7-binding activity, respectively (20,22).…”

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Structure and Mutagenesis Studies of the C-terminal Region of Licensing Factor Cdt1 Enable the Identification of Key Residues for Binding to Replicative Helicase Mcm Proteins

Jee

Mizuno

Kamada

et al. 2010

Journal of Biological Chemistry

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In eukaryotes, DNA replication is fired once in a single cell cycle before cell division starts to maintain stability of the genome. This event is tightly controlled by a series of proteins. Cdt1 is one of the licensing factors and is involved in recruiting replicative DNA helicase Mcm2-7 proteins into the pre-replicative complex together with Cdc6. In Cdt1, the C-terminal region serves as a binding site for Mcm2-7 proteins, although the details of these interactions remain largely unknown. Here, we report the structure of the region and the key residues for binding to Mcm proteins. We determined the solution structure of the C-terminal fragment, residues 450 -557, of mouse Cdt1 by NMR. The structure consists of a winged-helix domain and shows unexpected similarity to those of the C-terminal domain of Cdc6 and the central fragment of Cdt1, thereby implying functionalandevolutionaryrelationships.Structure-basedmutagenesis and an in vitro binding assay enabled us to pinpoint the region that interacts with Mcm proteins. Moreover, by performing in vitro binding and budding yeast viability experiments, we showed that ϳ45 residues located in the N-terminal direction of the structural region are equally crucial for recognizing Mcm proteins. Our data suggest the possibility that winged-helix domain plays a role as a common module to interact with replicative helicase in the DNA replication-licensing process.In eukaryotes, DNA replication is highly coordinated to retain the integrity of the genome. Whereas DNA replication in prokaryotes begins at a single site and stops at the end of the genome, eukaryotic genomes consist of multiple replication origins where DNA replication starts. These origins are synchronized so that they are activated only once in a single division cycle. A series of proteins, the origin recognition complex (ORC), 6 cell division cycle 6 homolog (Cdc6), chromatin licensing and DNA replication factor 1 (Cdt1), and minichromosome maintenance 2-7 (Mcm2-7) are known to play correlated roles in licensing (1-5). Oncogenic proliferation often causes abnormal expression of the proteins involved in the DNA-licensing process, thus emphasizing the importance of harmonious adjustments between these proteins (6). A complicated interaction network between these proteins has been reported (7), although the details at residue and atom levels remain largely unknown.Formation of a pre-replication complex at each origin is the first event in the replication process. ORC proteins bind initially to each replication origin of DNA. The DNA sequences of the origins where ORC binds have not been identified, except for those in Saccharomyces cerevisiae (8), suggesting that there may be other factors involved in addition to the sequences (9). The DNA strand at the origin needs to be unpaired to begin replication; therefore, the existence of replicative helicase is essential. Mcm2-7 proteins are believed to function as the replicative helicase in eukaryotes. Each Mcm2-7 protein consists of a conserved AAAϩ ATPase type C-termin...

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Section: Regions Of Winged-helix Domain For Binding To MCMsupporting

confidence: 86%

mentioning

confidence: 99%

Structure and Mutagenesis Studies of the C-terminal Region of Licensing Factor Cdt1 Enable the Identification of Key Residues for Binding to Replicative Helicase Mcm Proteins

Jee

Mizuno

Kamada

et al. 2010

Journal of Biological Chemistry

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“…These Cdc10 targets are transiently expressed during S phase (Lowndes et al, 1992;Kelly et al, 1993;Hofmann and Beach, 1994) and are under control of the DSC1 (DNA synthesis control) transcription factor (Lowndes et al, 1992;Hofmann and Beach, 1994;Baum et al, 1997). In asynchronous cells, IR induction levels of between approximately twofold (cdc22 ϩ ) and approximately fourfold (cdt2 ϩ ) after 160 min were observed ( Figure 3C) and contrasted markedly with IR-induction levels of between approximately sixfold (cdt2 ϩ ) and ϳ30-fold (cdc22 ϩ ) in irradiated G2-synchronized cells ( Figure 3D).…”

Section: Ir-induced Gene Expression In G2 Synchronized Cellsmentioning

confidence: 99%

Global Gene Expression Responses of Fission Yeast to Ionizing Radiation

Watson

Mata

Bähler

et al. 2004

MBoC

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A coordinated transcriptional response to DNA-damaging agents is required to maintain genome stability. We have examined the global gene expression responses of the fission yeast Schizosaccharomyces pombe to ionizing radiation (IR) by using DNA microarrays. We identified ϳ200 genes whose transcript levels were significantly altered at least twofold in response to 500 Gy of gamma IR in a temporally defined manner. The majority of induced genes were core environmental stress response genes, whereas the remaining genes define a transcriptional response to DNA damage in fission yeast. Surprisingly, few DNA repair and checkpoint genes were transcriptionally modulated in response to IR. We define a role for the stress-activated mitogen-activated protein kinase Sty1/Spc1 and the DNA damage checkpoint kinase Rad3 in regulating core environmental stress response genes and IR-specific response genes, both independently and in concert. These findings suggest a complex network of regulatory pathways coordinate gene expression responses to IR in eukaryotes. INTRODUCTIONExposure of cells to DNA-damaging agents such as ionizing radiation (IR) poses a significant threat to genome stability. Cells have therefore evolved complex response mechanisms to maintain genetic integrity after DNA damage. These include cell cycle delay, repair of DNA damage, transcriptional responses, and programmed cell death. Because disruption of any one of these DNA damage responses can lead to genomic instability and cancer, a comprehensive understanding of the individual components and their regulation is crucial.It is well established that many physiological responses to DNA damage are regulated at the level of gene expression. In Escherichia coli, the SOS response induces the expression of a network of genes after DNA damage through the regulatory LexA and RecA proteins (Friedberg et al., 1995). In lower eukaryotes, studies with the budding yeast Saccharomyces cerevisiae have identified a large number of damageinduced genes, including many involved in DNA metabolism and DNA repair (Aboussekhra et al., 1996;Gasch et al., 2001). In the distantly related fission yeast Schizosaccharomyces pombe, rhp51 ϩ , uvi15 ϩ , uvi31 ϩ , UVDE ϩ , and rhp16 ϩ have been shown to be DNA damage inducible (Bang et al., 1996;Davey et al., 1997;Shim et al., 2000).DNA damage checkpoint pathways function to delay the eukaryotic cell cycle in response to DNA damage, thus providing an opportunity for DNA repair. Central to the DNA damage checkpoint pathway are two highly conserved phosphoinositol-related kinases, Ataxia telangiectasia mutated (ATM) and ATM and Rad3-related (ATR) and their yeast homologs SpTel1 and SpRad3 in S. pombe, and ScTel1 and ScMec1 in S. cerevisiae. Activation of ATM and ATR kinases by DNA damage leads to cell cycle arrest through a number of downstream effector molecules including the effector kinases CHK1/SpChk1/ScChk1 and CHK2/SpCds1/ ScRad53 (for review, see Nyberg et al., 2002). In addition to regulating the cell cycle, DNA damage checkpoints also...

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“…CDC6 seems to cooperate with CDT1, a gene that was originally found in S. pombe (39) and is required for DNA replication in different organisms. (40)(41)(42)(43)(44)(45)(46) Cdt1 physically associates with Cdc6 (41) suggesting that it also forms part of the pre-RC.…”

Section: Introductionmentioning

confidence: 99%

Perpetuating the double helix: molecular machines at eukaryotic DNA replication origins

2003

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SummaryThe hardest part of replicating a genome is the beginning. The first step of DNA replication (called ''initiation'') mobilizes a large number of specialized proteins (''initiators'') that recognize specific sequences or structural motifs in the DNA, unwind the double helix, protect the exposed ssDNA, and recruit the enzymatic activities required for DNA synthesis, such as helicases, primases and polymerases. All of these components are orderly assembled before the first nucleotide can be incorporated. On the occasion of the 50th anniversary of the discovery of the DNA structure, we review our current knowledge of the molecular mechanisms that control initiation of DNA replication in eukaryotic cells, with particular emphasis on the recent identification of novel initiator proteins. We speculate how these initiators assemble molecular machines capable of performing specific biochemical tasks, such as loading a ringshaped helicase onto the DNA double helix.

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cdt1 is an essential target of the Cdc10/Sct1 transcription factor: requirement for DNA replication and inhibition of mitosis.

Cited by 223 publications

References 50 publications

Structure and Mutagenesis Studies of the C-terminal Region of Licensing Factor Cdt1 Enable the Identification of Key Residues for Binding to Replicative Helicase Mcm Proteins

Structure and Mutagenesis Studies of the C-terminal Region of Licensing Factor Cdt1 Enable the Identification of Key Residues for Binding to Replicative Helicase Mcm Proteins

Global Gene Expression Responses of Fission Yeast to Ionizing Radiation

Perpetuating the double helix: molecular machines at eukaryotic DNA replication origins

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