Plasmids Resembling 2- m DNA in the Osmotolerant Yeasts Saccharomyces bailii and Saccharomyces bisporus

Toh‐e, Akio; Araki, Hideki; Utatsu, Ikuyo; Oshima, Yasuji

doi:10.1099/00221287-130-10-2527

Cited by 29 publications

(41 citation statements)

References 10 publications

(3 reference statements)

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“…Several circular plasmids resembling the 2p plasmid of Saccharomyces cereuisiae have been described : pSRl , pSB3 and pSB4 from Zygosaccharomyces rouxii (Toh-e et al, 1982(Toh-e et al, , 1984, pSBl and pSB2 from 2. bailii (Toh-e et al, 1984), pSMl from 2. fermentatii (Utatsu et al, 1987) and pKD 1 from Kluyveromyces drosophilarurn . These cryptic plasmids, which we may call 2p-like plasmids, are found in high-copy-number in the host cell.…”

Section: Introductionmentioning

confidence: 99%

Characterization of a circular plasmid from the yeast Kluyveromyces waltii

Chen

Cong

Wésolowski-Louvel

et al. 1992

Journal of General Microbiology

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A new plasmid was found in the yeast Kluyoeromyces wdtii. This high-copy-number plasmid, named pKW1, is a double-stranded circular DNA plasmid of 5619 bp. It has several features characteristic of the 2ptype plasmids: presence of two inverted repeats and four open reading frames, as well as the interconversion of two isomeric forms. However, the nucleotide sequence shows little homology with known yeast plasmids. An ARS function was localized within a segment of 545 bp near one of the inverted repeats. Chimeric plasmids carrying this segment efficiently transformed K. wdtii. A strain of K. waltii cured of the plasmid (ciro) was also obtained. In the pKWl sequence, a functionally neutral region was found at which foreign DNA can be inserted with little effect on plasmid stability. Such constructions carrying the full sequence of pKWl replicated autonomously in a cir" host and were particularly stable. pK W 1-derived full-sequence plasmids also transformed K. thermotolerans, but not K. lactis.

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Section: Introductionmentioning

confidence: 99%

Characterization of a circular plasmid from the yeast Kluyveromyces waltii

Chen

Cong

Wésolowski-Louvel

et al. 1992

Journal of General Microbiology

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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.…”

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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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“…For unknown reasons, 2ptm DNA-like plasmids were found frequently in Zygosaccharomyces species. We have found five plasmids in Zygosaccharomyces yeasts: pSB1 and pSB2 from Z. bailii (23), pSR1 and pSB3 from Z. rouxii (23,24), and pSM1 from Z. fermentati (unpublished data). Recently, another 2,um DNA-like plasmid, pKD1, was discovered in Kluyveromyces drosophilarum (8).…”

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

et al. 1987

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The nucleotide sequence of two Zygosaccharomyces plasmids, pSB2 (5,415 base pairs), isolated from Zygosaccharomyces bailii, and pSMl (5,416 base pairs), isolated from Zygosaccharomycesfermentati Naganishi, was determined. The predicted amino acid sequences of open reading frames among six yeast plasmids that resemble 2,um DNA indicated regional sequence similarities among FLP proteins. Greater similarities were seen among Zygosaccharomyces plasmids (pSB2, pSB3, pSRl, and pSM1) than other combinations. A putative recognition site for the FLP enzyme of a Zygosaccharomyces plasmid also showed some conservation, especially in the 4 nucleotides flanking the central spacer region. From comparative studies of the sequences of putative genes of each plasmid, we propose an apparent phylogenetic relationship among yeast plasmids resembling 2,Lm DNA. Among the Zygosaccharomyces plasmids, pSB2 and pSRl are most closely related, since not only were the FLP enzymes of these two plasmids most closely related, but also the amino acid sequence of the putative P gene of pSRl showed clear homology with that of open reading frame B of pSB2.Since the discovery of 2,um DNA in Saccharomyces species (21; see reference 6 for a review), several yeast plasmids resembling the 2Rm plasmid were isolated and characterized. We call these plasmids 2,um DNA-like plasmids. For unknown reasons, 2ptm DNA-like plasmids were found frequently in Zygosaccharomyces species. We have found five plasmids in Zygosaccharomyces yeasts: pSB1 and pSB2 from Z. bailii (23), pSR1 and pSB3 from Z. rouxii (23,24), and pSM1 from Z. fermentati (unpublished data). Recently, another 2,um DNA-like plasmid, pKD1, was discovered in Kluyveromyces drosophilarum (8). Of these, the nucleotide sequences of 2,um DNA (13), pSR1 (2), pSB3 (25), and pKD1 (8) have been published. These plasmids, along with 2,um DNA from Saccharomyces cerevisiae, have several structural features in common: (i) they are closed circular DNA plasmids whose size is ca. 6 kilobases (kb), (ii) they have a pair of inverted repeats (IRs) consisting of several hundred bases, (iii) they are cryptic and exist in high copy number, and (iv) intramolecular recombination results in two isomeric forms. In spite of resemblances in their global structure, homology at the nucleotide sequence level is poor (23).Plasmid-encoded functions have been most extensively studied in 2,um DNA. The prominent features of genes on 2,um DNA are briefly summarized as follows. It has one replication origin, which is activated once in the cell cycle by the same replication mechanism as that for replication of chromosomes (16,28). It has four open reading frames (ORFs) or genes, A, B, C, and D. The A gene (or FLP gene) encodes a recombination enzyme which functions as a site-specific recombinase (7). The B and C genes produce trans-acting factors that are needed for the stable maintenance of this plasmid. These two genes, along with the cis-acting locus STB, participate in partitioning of plasmid molecules during cell division (14,15 Our...

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Plasmids Resembling 2- m DNA in the Osmotolerant Yeasts Saccharomyces bailii and Saccharomyces bisporus

Cited by 29 publications

References 10 publications

Characterization of a circular plasmid from the yeast Kluyveromyces waltii

Characterization of a circular plasmid from the yeast Kluyveromyces waltii

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

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

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