Elevated recombination rates in transcriptionally active DNA

Thomas, Barbara J.; Rothstein, Rodney

doi:10.1016/0092-8674(89)90584-9

Cited by 1,578 publications

(825 citation statements)

References 43 publications

Supporting

Mentioning

793

Contrasting

Unclassified

Order By: Relevance

“…An effect of transcription on genetic stability in a eukaryotic system was first observed using a segment of the RNA polymerase I transcribed ribosomal DNA locus -denoted HOT1 -that functioned as a cis-acting enhancer of recombination in Saccharomyces cerevisiae [2]. A similar increase in mitotic recombination and an additional elevation of spontaneous mutation rates in yeast has been associated with high levels of transcription carried out by RNA polymerase II [3][4][5][6][7]. The effect of such transcription-associated mutagenesis (TAM) and recombination (TAR) would be expected to be generally deleterious (e.g., the loss of a tumor suppressor), but may also be advantageous, promoting processes such as in immunoglobulin somatic hypermutation [8].…”

Section: Introductionmentioning

confidence: 99%

Transcription-associated mutagenesis in yeast is directly proportional to the level of gene expression and influenced by the direction of DNA replication

et al. 2007

View full text Add to dashboard Cite

A high level of transcription has been associated with elevated spontaneous mutation and recombination rates in eukaryotic organisms. To determine whether the transcription level is directly correlated with the degree of genomic instability, we have developed a tetracycline-regulated LYS2 reporter system to modulate the transcription level over a broad range in Saccharomyces cerevisiae. We find that spontaneous mutation rate is directly proportional to the transcription level, suggesting that movement of RNA polymerase through the target initiates a mutagenic process(es). Using this system, we also investigated two hypotheses that have been proposed to explain transcription-associated mutagenesis (TAM): 1) transcription impairs replication fork progression in a directional manner and 2) DNA lesions accumulate under high-transcription conditions. The effect of replication fork progression was probed by comparing the mutational rates and spectra in yeast strains with the reporter gene placed in two different orientations near a well-characterized replication origin. The effect of endogenous DNA damage accumulation was investigated by studying TAM in strains defective in nucleotide excision repair or in lesion bypass by the translesion polymerase Polζ. Our results suggest that both replication orientation and endogenous lesion accumulation play significant roles in TAM, particularly in terms of mutation spectra.

show abstract

Section: Introductionmentioning

confidence: 99%

Transcription-associated mutagenesis in yeast is directly proportional to the level of gene expression and influenced by the direction of DNA replication

et al. 2007

View full text Add to dashboard Cite

show abstract

“…Yeast C. versatilis NBR10650 (wild-type), S. cerevisiae YSH642 (Ansell et al, 1997), and W303-1A (Thomas and Rothstein, 1989;Wallis et al, 1989) were cultivated with shaking in YPAD medium (Watanabe et al, 2005) or Leu-dropout medium (Watanabe et al, 2006) after pre-cultivation for 3 days at 30 • C. To induce gene transcription, after precultivation in Leu-dropout medium containing 2% raffinose, pre-culture was added to Leu-dropout medium containing 1% raffinose and 1% galactose and cultivated with shaking at 30 • C. Escherichia coli JM109 (competent cells were purchased from Takara) was cultivated with shaking in LB medium (Sambrook and Russel, 2001) at 37 • C. Culture plates were prepared by solidifying liquid medium through the addition of 2% w/v agar and were incubated at 30 • C or 37 • C.…”

Section: Strains and Mediamentioning

confidence: 99%

Expression of glycerol 3‐phosphate dehydrogenase gene (CvGPD1) in salt‐tolerant yeast Candida versatilis is stimulated by high concentrations of NaCl

Watanabe¹,

Nagayama²,

Tamai³

2007

Yeast

View full text Add to dashboard Cite

We cloned the glycerol 3-phosphate dehydrogenase (GPDH) gene (CvGPD1 ) from salttolerant yeast Candida versatilis. When CvGPD1 was expressed in glycerol synthesisdeficient Saccharomyces cerevisiae cells, the salt tolerance of the recombinant strain was enhanced, and NADP + -dependent GPDH (EC 1.1.1.94), Cvgpd1p synthesis and recovery of glycerol synthesis were confirmed. The transcription of CvGPD1 in C. versatilis cells was stimulated by high concentrations of NaCl. The relationship between expression of CvGPD1 and growth of C. versatilis cells in the mash of Japanese seasonings (miso-and shoyu-moromi ) is also discussed. The Accession No. for the sequenced DNA fragment is AB296385.

show abstract

“…The following yeast strains were used: Zygosaccharomyces rouxii NRRL 2547 (obtained from C. Kurtzman), Saccharomyces cerevisiae W303-1A (MATα leu2-3/112 ura3-1 trp1-1 his3-11/15 ade2-1 can1-100 GAL SUC 2 mal 0; Thomas and Rothstein, 1989) and S. cerevisiae yJN009-1D (W303-1A dak1 :: KAN-MX4 dak2 :: HIS3-MX6; Molin M, Norbeck J, Blomberg A, J. Biol. Chem.…”

Section: Microorganisms Plasmids and Culture Conditionsmentioning

confidence: 99%

Cloning, sequencing and characterization of a gene encoding dihydroxyacetone kinase from Zygosaccharomyces rouxii NRRL2547

Wang

Kayingo

Blomberg

et al. 2002

Yeast

View full text Add to dashboard Cite

The dihydroxyacetone pathway, an alternative pathway for the dissimilation of glycerol via reduction by glycerol dehydrogenase and subsequent phosphorylation by dihydroxyacetone (DHA) kinase, is activated in the yeasts Saccharomyces cerevisiae and Zygosaccharomyces rouxii during osmotic stress. In experiments aimed at investigating the physiological function of the DHA pathway in Z. rouxii, a typical osmotolerant yeast, we cloned and characterized a DAK gene encoding dihydroxyacetone kinase from Z. rouxii NRRL 2547. Sequence analysis revealed a 1761 bp open reading frame, encoding a peptide composed of 587 deduced amino acids with the predicted molecular weight of 61 664 Da. As the amino acid sequence was most closely homologous (68% identity) to the S. cerevisiae Dak1p, we named the gene and protein ZrDAK1 and ZrDak1p, respectively. A putative ATP binding site was also found but no consensus element associated with osmoregulation was found in the upstream region of the ZrDAK1 gene. The ZrDAK1 gene complemented a S. cerevisiae W303-1A dak1 dak2 strain by improving the growth of the mutant on 50 mmol/l dihydroxyacetone and by increasing the tolerance to dihydroxyacetone in a medium containing 5% sodium chloride, suggesting that it is a functional homologue of the S. cerevisiae DAK1. However, expression of the ZrDAK1 gene in the S. cerevisiae dak1 dak2 strain had no significant effect on glycerol levels during osmotic stress. The ZrDAK1 sequence has been deposited in the public data bases under Accession No. AJ294719; regions upstream and downstream of ZrDAK1 are deposited as Accession Nos AJ294739 and AJ294720, respectively.

show abstract

Elevated recombination rates in transcriptionally active DNA

Cited by 1,578 publications

References 43 publications

Transcription-associated mutagenesis in yeast is directly proportional to the level of gene expression and influenced by the direction of DNA replication

Transcription-associated mutagenesis in yeast is directly proportional to the level of gene expression and influenced by the direction of DNA replication

Expression of glycerol 3‐phosphate dehydrogenase gene (CvGPD1) in salt‐tolerant yeast Candida versatilis is stimulated by high concentrations of NaCl

Cloning, sequencing and characterization of a gene encoding dihydroxyacetone kinase from Zygosaccharomyces rouxii NRRL2547

Contact Info

Product

Resources

About