Yeast plasmids resembling 2 micron DNA: regional similarities and diversities at the molecular level

Utatsu, Ikuyo; Sakamoto, Susumu; Imura, Toshiaki; Toh‐e, Akio

doi:10.1128/jb.169.12.5537-5545.1987

Cited by 77 publications

(58 citation statements)

References 27 publications

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“…We found that 2 DNA was present in 38 of the strains examined, including several S. bayanus and S. paradoxus isolates. Plasmids resembling 2 DNA have been described in even more distant relatives of S. cerevisiae (36).…”

Section: Resultsmentioning

confidence: 99%

Yeast prions [URE3] and [PSI ⁺ ] are diseases

Nakayashiki

Kurtzman

Edskes

et al. 2005

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

236

232

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Section: Resultsmentioning

confidence: 99%

Yeast prions [URE3] and [PSI ⁺ ] are diseases

Nakayashiki

Kurtzman

Edskes

et al. 2005

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

236

232

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“…We tested a subset of these predictions by site-directed mutagenesis of the Rep1 protein. The choice of Rep1p over Rep2p for this analysis was dictated by the higher degree of amino acid conservation for the former among yeast plasmids that resemble the 2m circle in structural and functional organization (27). Mutations were targeted to invariant or highly conserved amino acids.…”

Section: Resultsmentioning

confidence: 99%

Mutations in a Partitioning Protein and Altered Chromatin Structure at the Partitioning Locus Prevent Cohesin Recruitment by the Saccharomyces cerevisiae Plasmid and Cause Plasmid Missegregation

Yang

Mehta

Uzri

et al. 2004

Molecular and Cellular Biology

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The 2m circle is a highly persistent "selfish" DNA element resident in the Saccharomyces cerevisiae nucleus whose stability approaches that of the chromosomes. The plasmid partitioning system, consisting of two plasmid-encoded proteins, Rep1p and Rep2p, and a cis-acting locus, STB, apparently feeds into the chromosome segregation pathway. The Rep proteins assist the recruitment of the yeast cohesin complex to STB during the S phase, presumably to apportion the replicated plasmid molecules equally to daughter cells. The DNAprotein and protein-protein interactions of the partitioning system, as well as the chromatin organization at STB, are important for cohesin recruitment. Rep1p variants that are incompetent in binding to Rep2p, STB, or both fail to assist the assembly of the cohesin complex at STB and are nonfunctional in plasmid maintenance. Preventing the cohesin-STB association without impeding Rep1p-Rep2p-STB interactions also causes plasmid missegregation. During the yeast cell cycle, the Rep1p and Rep2p proteins are expelled from STB during a short interval between the late G 1 and early S phases. This dissociation and reassociation event ensures that cohesin loading at STB is replication dependent and is coordinated with chromosomal cohesin recruitment. In an rsc2⌬ yeast strain lacking a specific chromatin remodeling complex and exhibiting a high degree of plasmid loss, neither Rep1p nor the cohesin complex can be recruited to STB. The phenotypes of the Rep1p mutations and of the rsc2⌬ mutant are consistent with the role of cohesin in plasmid partitioning being analogous to that in chromosome partitioning.The 2m circle of Saccharomyces cerevisiae is a high-copynumber selfish extrachromosomal DNA element that resides in the nucleus and propagates itself stably in the cell population (2, 29). The plasmid does not seem to confer any selective advantage to its host under normal laboratory growth conditions, nor does it pose any noticeable disadvantage as long as the copy number does not rise significantly above the steadystate value of approximately 60 per cell. The stability of the plasmid approaches that of the yeast chromosomes, with the loss rate being as low as 10 Ϫ4 to 10 Ϫ5 per cell per generation. The structural organization of the plasmid and its genetic potential are devoted to two goals: (i) efficient plasmid segregation during cell division and (ii) the maintenance of the plasmid copy number with only modest deviations from the mean.The presence of a typical yeast replication origin in its sequence permits each plasmid molecule to be replicated once per cell cycle (35). A stability system consisting of two plasmidencoded proteins (Rep1p and Rep2p) and a cis-acting partitioning locus (STB) mediates equal or nearly equal distribution of the replicated molecules into daughter cells. The direct observation of reporter plasmids tagged with fluorescence indicates that the plasmid molecules are organized into a tightknit cluster in the nucleus and segregate as a cluster. Hence, the copy number relevant ...

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“…34). In particular, they appear to encode site-specific recombinases related to FLP and undergo high-frequency intramolecular recombination between inverted repeat domains both in their native species and in S. cerevisiae (32,(34)(35)(36)(37)(38)(39). However, only the R recombinase has previously been shown to work in a multicellular organism (33), and none has been used in an animal genome.…”

Section: And Plants (24)mentioning

confidence: 99%

Multiple new site-specific recombinases for use in manipulating animal genomes

Nern

Pfeiffer

Svoboda

et al. 2011

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

159

171

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Site-specific recombinases have been used for two decades to manipulate the structure of animal genomes in highly predictable ways and have become major research tools. However, the small number of recombinases demonstrated to have distinct specificities, low toxicity, and sufficient activity to drive reactions to completion in animals has been a limitation. In this report we show that four recombinases derived from yeast-KD, B2, B3, and R-are highly active and nontoxic in Drosophila and that KD, B2, B3, and the widely used FLP recombinase have distinct target specificities. We also show that the KD and B3 recombinases are active in mice.gene expression | genetic engineering S ite-specific DNA recombinases are widely used in multicellular organisms to manipulate the structure of genomes and, in turn, to control gene expression (for reviews see refs. 1-4). These enzymes, derived from bacteria and fungi, catalyze directionally sensitive DNA exchange reactions between short (30-40 nucleotides) target site sequences that are specific to each recombinase (5). These reactions enable four basic functional modules-excision/insertion, inversion, translocation and cassette exchange-that have been used individually or combined in a wide range of configurations to control gene expression (Fig. 1A).The use of site-specific recombination to manipulate genomes has been limited by the availability of recombinases with high activity, distinct site specificity, and low toxicity. In Drosophila, the most widely used recombinase is FLP, encoded by the Saccharomyces cerevisiae 2-μm plasmid (6). FLP was first shown to work in a heterologous, multicellular organism by Golic and Lindquist in 1989 (7) who demonstrated the excision reaction on chromosomally inserted target sites (FRTs). Since that time FLP/FRT recombination has been widely used in Drosophila in applications based on excision (8) and translocation (9-11).Complex manipulations of genome structure can require the use of more than one of the modules diagrammed in Fig. 1A, or parallel independent implementations of the same module, in a single individual. To accomplish such manipulations, the modules must be implemented with different recombinases that do not recognize each other's target sites. Similarly, a number of powerful methods have been developed for using the excision and inversion reactions to control expression of a transgene specifically in cells where two independent gene expression patterns overlap (2,3,12,13). Such intersectional methods rely on pairs of orthogonal recombinases; see, for example, Fig. 1B. For these reasons, we sought to discover additional recombinases with distinct site-specificity.FLP recombinase has been mutated to recognize altered FRT sites, but some cross-reaction still remains (14, 15). Cre, encoded by the bacteriophage P1, is the most widely used recombinase in mammalian cells (16)(17)(18). Cre functions in Drosophila (19), but exhibits obvious toxicity (20), a problem also observed in mammalian cells (21, 22; reviewed in ref. 23) an...

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Yeast plasmids resembling 2 micron DNA: regional similarities and diversities at the molecular level

Cited by 77 publications

References 27 publications

Yeast prions [URE3] and [PSI ⁺ ] are diseases

Yeast prions [URE3] and [PSI ⁺ ] are diseases

Mutations in a Partitioning Protein and Altered Chromatin Structure at the Partitioning Locus Prevent Cohesin Recruitment by the Saccharomyces cerevisiae Plasmid and Cause Plasmid Missegregation

Multiple new site-specific recombinases for use in manipulating animal genomes

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Yeast plasmids resembling 2 micron DNA: regional similarities and diversities at the molecular level

Cited by 77 publications

References 27 publications

Yeast prions [URE3] and [PSI + ] are diseases

Yeast prions [URE3] and [PSI + ] are diseases

Mutations in a Partitioning Protein and Altered Chromatin Structure at the Partitioning Locus Prevent Cohesin Recruitment by the Saccharomyces cerevisiae Plasmid and Cause Plasmid Missegregation

Multiple new site-specific recombinases for use in manipulating animal genomes

Contact Info

Product

Resources

About

Yeast prions [URE3] and [PSI ⁺ ] are diseases

Yeast prions [URE3] and [PSI ⁺ ] are diseases