Switching genes in Schizosaccharomyces pombe

Gütz, Herbert; Schmidt, Henning

doi:10.1007/bf00421601

Cited by 77 publications

(38 citation statements)

References 14 publications

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“…As a result of a search by BLAST in the databases of the NCBI, the swi3 ORF showed high similarity in amino acid sequence with mammalian and mouse timeless-interacting protein (Tipin) (2e-09 and 8e-08, respectively). The swi3-146, swi3-157, and swi3-1 mutations, causing defects in mating type switching (22), were identified by direct sequencing of PCR-amplified mutant alleles from strains E146, E157, and SG381, respectively. The swi3-146, swi3-157, and swi3-1 mutations changed the third [G to A (M1I)] and the 259th nucleotides [C to T(ochre)] and inserted A after the 132nd nucleotide (frame shift) of the wild-type swi3 ORF, respectively (Fig.…”

Section: Resultsmentioning

confidence: 99%

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Biochemical Interactions between Proteins and mat1 cis-Acting Sequences Required for Imprinting in Fission Yeast

Lee

Grewal

Klar

2004

Molecular and Cellular Biology

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DNA recombination required for mating type (mat1) switching in Schizosaccharomyces pombe is initiated by mat1 imprinting. The imprinting event is regulated by mat1 cis-acting elements and by several trans-acting factors, including swi1 (for switch), swi3, swi7, and sap1. swi1 and swi3 were previously shown to function in dictating unidirectional mat1 DNA replication by controlling replication fork movement around the mat1 region and, second, by pausing fork progression around the imprint site. With biochemical studies, we investigated whether the trans-acting factors function indirectly or directly by binding to the mat1 cis-acting sequences. First, we report the identification and DNA sequence of the swi3 gene. swi3 is not essential for viability, and, like the other factors, it exerts a stimulatory effect on imprinting. Second, we showed that only Swi1p and Swi3p interact to form a multiprotein complex and that complex formation did not require their binding to a DNA region defined by the smt-0 mutation. Third, we found that the Swi1p-Swi3p complex physically binds to a region around the imprint site where pausing of replication occurs. Fourth, the protein complex also interacted with the mat1-proximal polar terminator of replication (RTS1). These results suggest that the stimulatory effect of swi1 and swi3 on switching and imprinting occurs through interaction of the Swi1p-Swi3p complex with the mat1 regions.The fission yeast Schizosaccharomyces pombe is a haploid eukaryotic microorganism whose cells exhibit two different mating types, called P (plus) and M (minus) (18,33). The mating type is determined by the genetic information in the mat1 locus, which contains either the mat1M or the mat1P allele, each of which encodes two transcripts essential for controlling cell type (24). A copy of the mating type genetic information is also present at the mat2P and mat3M"donor" loci that are located centromere-distal to mat1 and are transcriptionally silent. The mating type switches when a copy of either the mat2P or mat3M DNA transposes and substitutes for mat1 by DNA recombination, where the transposed genetic information is expressed (6,12,26).Switching occurs spontaneously at a high frequency in "homothallic" strains (h 90 , i.e., homothallic, 90% sporulation) during mitotic growth, and it follows an interesting nonrandom pattern within a cell lineage (17,28,34). Division of an unswitchable parental cell produces two daughter cells, one of which is programmed to be switching competent and the other of which is not. After cell division of a switching-competent cell, a switched and a switching-competent daughter cell are generated. On the other hand, a switching-noncompetent cell undergoes a cell division to produce a switching-competent and a noncompetent cell. Therefore, this switching pattern in the cell lineage results in only one cell switching among four granddaughter cells after two consecutive asymmetrical cell divisions of an unswitchable cell. These rules of so-called onein-four and consecutive switching a...

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Section: Resultsmentioning

confidence: 99%

“…In earlier research, genetic screening for mutations affecting both mating type switching and the level of double-strand break implicated swi1, swi3, and swi7 in the switching process (16,22). Doublestrand break has also been shown to efficiently initiate meiotic mat1 gene conversion (30).…”

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confidence: 99%

Biochemical Interactions between Proteins and mat1 cis-Acting Sequences Required for Imprinting in Fission Yeast

Lee

Grewal

Klar

2004

Molecular and Cellular Biology

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“…The primers used in these procedures are available on request. Mutations and epitopetagged genes have previously been described for cdc25-22 (Fantes 1979), swi1D (swi1TKan r ) , swi3D (swi3TKan r ) (Noguchi et al 2004), swi3-1 (Gutz and Schmidt 1985), cds1D (cds1Tura4 1 ) (Boddy et al 1998) , swi1-GFP (swi1-GFP-Kan r ) , swi3-GFP (swi3-GFPKan r ) (Noguchi et al 2004), swi1-3FLAG (swi1-3FLAG-Kan r ) (Noguchi et al 2004), uve1D (uve1TLEU2) (Yonemasu et al 1997), rad13D (rad13Tura4 1 ) (Yonemasuet al 1997), and orp1-5FLAG (orp1-5FLAG-Kan r ) (Ogawa et al 1999).…”

Section: General Techniquesmentioning

confidence: 99%

Sap1 Promotes the Association of the Replication Fork Protection Complex With Chromatin and Is Involved in the Replication Checkpoint in Schizosaccharomyces pombe

Noguchi

2007

Genetics

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Sap1 is involved in replication fork pausing at rDNA repeats and functions during mating-type switching in Schizosaccharomyces pombe. These two roles are dependent on the ability of Sap1 to bind specific DNA sequences at the rDNA and mating-type loci, respectively. In S. pombe, Swi1 and Swi3 form the replication fork protection complex (FPC) and play important roles in the activation of the replication checkpoint and the stabilization of stalled replication forks. Here we describe the roles of Sap1 in the replication checkpoint. We show that Sap1 is involved in the activation of the replication checkpoint kinase Cds1 and that sap1 mutant cells accumulate spontaneous DNA damage during the S-and G2-phases, which is indicative of fork damage. We also show that sap1 mutants have a defect in the resumption of DNA replication after fork arrest. Sap1 is localized at the replication origin ori2004 and this localization is required for the association of the FPC with chromatin. We propose that Sap1 is required to recruit the FPC to chromatin, thereby contributing to the activation of the replication checkpoint and the stabilization of replication forks.

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“…By 20.2% identity in a region of 104 amino acids. Rad5O is required for recombination (mitotic and meiotic) and for DSB DNA repair during vegetative growth in S. cerevisiae (18,22).…”

Section: Discussionmentioning

confidence: 99%

“…The first class is composed of three trans-acting switch (swi) loci (swil, swi3, and swi7), which are required for producing a high level of DSB (6,14,20). We have recently shown that swi7 encodes the catalytic subunit of DNA polymerase a (45).…”

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Sap1, a protein that binds to sequences required for mating-type switching, is essential for viability in Schizosaccharomyces pombe.

1994

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The pattern of mating-type switching in cell pedigrees of the fission yeast Schizosaccharomyces pombe is dictated by the inheritance of specific DNA chains at the mating-type locus (mat1). The recombination event essential for switching is initiated by a site-specific double-strand break at mat). The switch-activating protein, Sapl, binds in vitro to a mat] cis-acting site that was shown earlier to be essential for efficient mating-type switching. We isolated the sap) gene by using oligonucleotides corresponding to the amino acid sequence of purified Sapl protein. (12,32). Consequently, a colony grown from a single cell of either P or M mating type contains nearly equal proportions of P and M cells (for reviews, see references 3, 13, 21, and 27). Pedigree analysis of single cells grown under starvation conditions, in which mating and sporulation are induced, revealed that switching is not random but follows several rules. First, the mating-type switching is formally a G2 event, as only one of the two daughters of a switching-competent cell actually switches. Second, two consecutive developmentally asymmetric cell divisions are required to restrict switching to only one cell among four related sister and cousin cells. The first asymmetric division of a switching-incompetent cell produces two unswitched cells that differ in their potential for switching: one is switching competent, while the other, like its parent, is not. The second asymmetric division of the switching competent cell produces one switched cell and one unswitched cell (35). Third, the switching potential is inherited by the sister of the recently switched cell, which can repeat the process of switching again and again, producing a chain of consecutive switching (15,26). The observed pattern of switching is analogous to that of the mammalian stem cell lineage, because the parental cell

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Switching genes in Schizosaccharomyces pombe

Cited by 77 publications

References 14 publications

Biochemical Interactions between Proteins and mat1 cis-Acting Sequences Required for Imprinting in Fission Yeast

Biochemical Interactions between Proteins and mat1 cis-Acting Sequences Required for Imprinting in Fission Yeast

Sap1 Promotes the Association of the Replication Fork Protection Complex With Chromatin and Is Involved in the Replication Checkpoint in Schizosaccharomyces pombe

Sap1, a protein that binds to sequences required for mating-type switching, is essential for viability in Schizosaccharomyces pombe.

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