Telomere Sequences Attached to Nuclearly Migrated Yeast Linear Plasmid

Takata, Hideki; Fukuda, Kei; Meinhardt, Friedhelm; Gunge, Norio

doi:10.1006/plas.1999.1454

Cited by 5 publications

(11 citation statements)

References 27 publications

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“…Although the added telomeric sequences varied in the arrangement of TG 1±3 repeats among the individual pTLU plasmids isolated, those at the two ends of any given pTLUs were identical. To explain this ®nding, we proposed a two-step model of telomere addition at the plasmid ends, based on a racketframe model involving, ®rst, telomerase action on one end, and then a strand invasion-copying mechanism on the opposite end (Sakaguchi 1990;Takata et al 2000).…”

Section: Discussionmentioning

confidence: 99%

“…cerevisiae strains harboring pTLU or pRLU1 were used: K12-PT2 (MATa leu2 ura3 rho + pTLU2), K12-PT3 (MATa leu2 ura3 rho + pTLU2), K12-PT3 (MATa leu2 ura3 rho + pTLU3), XS95-6C-PT101 (MATa rad52 his3 leu2 ura3 trpl can r rho + pTLU101) and K12-U2-S1 (MATa leu2 ura3 rho + pRLU1). K12-PT2, K12-PT3 and K12-U2-S1 are isogenic and were derived from K12-PC (MATa leu2 ura3 rho + pGKL2 pCLU1) (Gunge et al 1995;Takata et al 2000). The circular plasmid pRLU1 (8372 bp) arose spontaneously from pCLU1 (8442 bp) by the loss of a 70-bp stretch from an ITR-terminus through a homologous recombination between short direct repeats within the ITR ends .…”

Section: Yeast Strains and Plasmidsmentioning

confidence: 99%

“…The nucleotide sequence between positions 6 and 23 is identical to that of pTLU2 and was omitted for simplicity. Sequence data for pTLU2 are taken from Takata et al (2000). In this study, the sequences of the pTLU2 derivatives were determined for about 100 bp around the ITR, but the alteration presumably continues toward the terminus of the added telomere, which is indicated by the broken lines (see text) on SC-Ura, which carried pTLU2 copies with the original telomeric sequence, was plated on SC for single colonies.…”

Section: Rearrangement Of Telomeric Repeat Sequencesmentioning

confidence: 99%

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Progressive alteration of telomeric sequences at one end of a yeast linear plasmid and its possible association with reduced plasmid stability

Takata

Gunge

2001

Mol Gen Genomics

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After selection for migration into the nucleus, a cytoplasmic yeast linear plasmid bearing an inverted terminal repeat (ITRs) at each end replicates in Saccharomyces cerevisiae in a linear form, called pTLU, which carries host telomeric repeats (TG(1-3))(n) of about 300-350 bp added to the ITR ends. We previously showed that the nucleotide composition of the added telomeric sequences varied among individual pTLU isolates, while those on the two ends of any given pTLU were always identical. The telomeric sequences of pTLU remained unchanged over numbers of cell generations when cells were selected for expression of the plasmid-borne nuclear marker. We report here that progressive alterations in telomeric sequences can be detected in cells which are grown under non-selective conditions. Surprisingly, in any given molecule, the telomeric alterations occur exclusively on one side, either the left or the right end, while the sequence at the opposite end remained identical to the original, suggesting a difference in the mode of DNA replication between the plasmid ends. These alterations occur over a broad area extending from the termini of telomeres to nucleotides near the junction between the telomeric sequences and the pTLU-ITR, implying that the plasmid ends undergo successive rounds of extension and contraction. Clonal analysis under non-selective conditions indicated that the alterations in telomeric sequences are generally associated with extreme instability of the pTLU plasmid.

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Section: Discussionmentioning

confidence: 99%

Section: Yeast Strains and Plasmidsmentioning

confidence: 99%

Section: Rearrangement Of Telomeric Repeat Sequencesmentioning

confidence: 99%

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Progressive alteration of telomeric sequences at one end of a yeast linear plasmid and its possible association with reduced plasmid stability

Takata

Gunge

2001

Mol Gen Genomics

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“…This ®nding contrasts with the analysis in S. cerevisiae, where telomere-associated linear plasmids were frequently observed . We presume that the ITR ends of pCLU1 can be used as a substrate by the telomerase of S. cerevisiae (Takata et al 2000) but not by the K. lactis telomerase.…”

Section: Intergeneric Transfer Of Pclu1mentioning

confidence: 99%

“…Based on the phenotypes conferred by these markers, plasmid migration from the cytoplasm into the nucleus could be monitored easily. Using S. cerevisiae, we studied the genetic modi®cations present in the nuclear forms of pCLU1 and demonstrated how the telomere sequences are added to the ITRs at the ends of the plasmid (Takata et al 2000).…”

Section: Introductionmentioning

confidence: 99%

Relocation of a cytoplasmic yeast linear plasmid to the nucleus is associated with circularization via nonhomologous recombination involving inverted terminal repeats

et al. 2000

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The linear plasmid pCLU1 from the yeast Kluyveromyces lactis normally replicates in the cytoplasm, with the aid of the helper linear plasmid pGKL2, using terminal protein (TP) as a primer. However, it relocates to the nucleus when selection is applied for the expression of a plasmid-borne nuclear marker. Migration to the nucleus occurred in K. lactis at a frequency of about 10(-3)/cell ten or more times higher than the rate observed in Saccharomyces cerevisiae. The nuclear plasmids existed only in a circularized form in K. lactis, while in S. cerevisiae a telomere-associated linear form is also found. Sequence analysis showed that circularization in K. lactis was caused by non-homologous recombination between the inverted terminal repeat (ITR) at the ends of the linear form and non-specific internal target sites in pCLU1. No sequence similarity existed among the junction sites, indicating that the free ITR end plays a crucial role in circularization. In S. cerevisiae, circular plasmids were generated not only by nonhomologous recombination, but also by homologous recombination between short direct repeats within pCLU1. Circularization via the ITR end was observed independently of RAD52 activity. Sequences highly homologous to ARS core elements, 5'-ATTTATTGTTTT-3' for K. lactis and 5'-(A/T)TTTAT(T/G)TTT(A/T)-3' for S. cerevisiae, were detected at multiple sites in the nuclear forms of the plasmids.

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Linear DNA Plasmids and Killer System of Kluyveromyces lactis

Gunge

Tokunaga

2004

Genetics and Biotechnology

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Telomere Sequences Attached to Nuclearly Migrated Yeast Linear Plasmid

Cited by 5 publications

References 27 publications

Progressive alteration of telomeric sequences at one end of a yeast linear plasmid and its possible association with reduced plasmid stability

Progressive alteration of telomeric sequences at one end of a yeast linear plasmid and its possible association with reduced plasmid stability

Relocation of a cytoplasmic yeast linear plasmid to the nucleus is associated with circularization via nonhomologous recombination involving inverted terminal repeats

Linear DNA Plasmids and Killer System of Kluyveromyces lactis

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