TY - JOUR AB - Transcriptional reinduction memory is a phenomenon whereby cells "remember" their transcriptional response to a previous stimulus such that subsequent encounters with the same stimulus can result in altered gene expression kinetics. Chromatin structure is thought to play a role in certain transcriptional memory mechanisms, leading to questions as to whether and how memory can be actively maintained and inherited to progeny through cell division. Here we summarize efforts towards dissecting chromatin-based transcriptional memory inheritance ofGALgenes inSaccharomyces cerevisiae. We focus on methods and analyses ofGAL(as well asMALandINO) memory in single cells and discuss the challenges in unraveling the underlying mechanisms in yeast and higher eukaryotes. AU - Bheda, P. AU - Kirmizis, A.* AU - Schneider, R. C1 - 59707 C2 - 49018 CY - One New York Plaza, Suite 4600, New York, Ny, United States SP - 1029–1035 TI - The past determines the future: Sugar source history and transcriptional memory. JO - Curr. Genet. VL - 66 PB - Springer PY - 2020 SN - 0172-8083 ER - TY - JOUR AB - An episomal DNA vector (YpJA18), encoding two selectable recombinant yeast genes (TRP1, URA3), was constructed to assess the fidelity of DNA repair in haploid repair-competent (RAD) wild-type yeast and several radiation-sensitive mutants. Either a DNA double-strand break (DSB) or a double-strand gap of 169 bp (DSG) was introduced by restriction enzymes in-vitro within the coding sequence of the URA3 gene of this vector. To eliminate transfer artefacts, selection was first applied for the undamaged TRP1 gene followed by counter selection for URA3 gene activity, which indicated correct repair of the DSB and DSG. Correct repair of the damaged URA3 gene was found to be about 90% in RAD cells (normalized for the expression of undamaged URA3 in TRP + transformants). Plasmids isolated from the transformants (URA + TRP +) carry both unique sites (ApaI and NcoI) within the URA3 gene indicating the precise restitution of the 169-bp gap. An excision-repair-defective rad4-4 mutant repaired these lesions as correctly as RAD cells, whereas the mutants rad50-1, rad51-1 and rad54-1, proven to be defective in DSB repair and mitotic recombination, showed less than 5% correct repair of such lesions. In contrast, a representative of the RAD6 epistasis group of genes, the rev2-1 mutant which is sensitive towards UV and ionizing radiation, had a significantly reduced ability (about 20%) for the correct repair of both DSBs and DSGs. AU - Jha, B. AU - Ahne, F. AU - Eckardt-Schupp, F. C1 - 20145 C2 - 13321 SP - 402-407 TI - The Use of Double Marker Shuttle Vector to Study DNA Double Strand Break Repair in Wild- Type and Radiation-Sensitive Mutants of the Yeast Saccharomyces Cerevisiae. JO - Curr. Genet. VL - 23 IS - 5 PY - 1993 SN - 0172-8083 ER - TY - JOUR AB - An episomal DNA vector (YpJA18), encoding two selectable recombinant yeast genes (TRP1, URA3), was constructed to assess the fidelity of DNA repair in haploid repair-competent (RAD) wild-type yeast and several radiation-sensitive mutants. Either a DNA double-strand break (DSB) or a double-strand gap of 169 bp (DSG) was introduced by restriction enzymes in-vitro within the coding sequence of the URA3 gene of this vector. To eliminate transfer artefacts, selection was first applied for the undamaged TRP1 gene followed by counter selection for URA3 gene activity, which indicated correct repair of the DSB and DSG. Correct repair of the damaged URA3 gene was found to be about 90 % in RAD cells (normalized for the expression of undamaged URA3 in TRP+ transformants). Plasmids isolated from the transformants (URA+ TRP+) carry both unique sites (ApaI and NcoI) within the URA3 gene indicating the precise restitution of the 169-bp gap. An excision-repair-defective rad4-4 mutant repaired these lesions as correctly as RAD cells, whereas the mutants rad50-1, rad51-1 and rad54-1, proven to be defective in DSB repair and mitotic recombination, showed less than 5% correct repair of such lesions. In contrast, a representative of the RAD6 epistasis group of genes, the rev2-1 mutant which is sensitive towards UV and ionizing radiation, had a significantly reduced ability (about 20 %) for the correct repair of both DSBs and DSGs. AU - Jha, B.S. AU - Ahne, F. AU - Eckardt-Schupp, F. C1 - 40436 C2 - 40038 SP - 402-407 TI - The use of a double-marker shuttle vector to study DNA double-strand break repair in wild-type and radiation-sensitive mutants of the yeast Saccharomyces cerevisiae. JO - Curr. Genet. VL - 23 IS - 5-6 PY - 1993 SN - 0172-8083 ER - TY - JOUR AB - The REV2 gene of Saccharomyces cerevisiae was cloned and sequenced; it contains an open reading frame of 1986 bp with a coding potential of 662 amino acids. Interruption of the chromosomal REV2 gene by integrating the URA3 gene coupled with partial deletion of the 3' terminal region produced viable haploid rev2Δ mutants. This indicates that the REV2 gene is non-essential for growth. The rev2Δ mutant is slightly more UV-sensitive than strains carrying various rev2 alleles (rev2-1, rev2x, rad5-1, rad5-8). The putative Rev2 protein is probably a globular protein containing a highly conserved nucleotide-binding site and two zinc-finger domains. AU - Ahne, F. AU - Baur, M. AU - Eckardt-Schupp, F. C1 - 40513 C2 - 38797 SP - 277-282 TI - The REV2 gene of Saccharomyces cerevisiae: Cloning and DNA sequence. JO - Curr. Genet. VL - 22 IS - 4 PY - 1992 SN - 0172-8083 ER - TY - JOUR AB - Repair under non-growth conditions of DNA double-strand breaks (DSB) and chromatin sites sensitive to S1 endonuclease (SSS) induced by 60Cobalt-gamma rays were monitored in repair-competent and deficient strains of Saccharomyces cerevisiae by pulsed field gel-electrophoresis. In stationary-phase cells of a repair-competent RAD diploid, and an excision-deficient rad3-2 diploid, SSS are repaired as efficiently as DSB, whereas in a repair-competent RAD haploid, and a rad 50-1 diploid, neither SSS nor DSB are repaired. The rad18-2 diploid repairs DSB well but is defective in SSS repair. Obviously, SSS repair in yeast chromatin, like DSB repair, depends on recombination, but unlike DSB repair depends additionally on RAD18 function. AU - Geigl, E.M. AU - Eckardt-Schupp, F. C1 - 40757 C2 - 38921 SP - 33-37 TI - Repair of gamma ray-induced S1 nuclease hypersensitive sites in yeast depends of homologous mitotic recombination and a RAD18-dependent function. JO - Curr. Genet. VL - 20 IS - 1-2 PY - 1991 SN - 0172-8083 ER - TY - JOUR AB - The RAD4 gene of yeast required for the incision step of DNA excision repair and the REV2 (= RAD5) gene involved in mutagenic DNA repair could not be isolated from genomic libraries propagated in E. coli regardless of copy number of the shuttle vector in yeast. Transformants with plasmids conferring UV resistance to a rad4-4 or a rev2-1 mutant were only recovered if yeast was transformed directly without previous amplification of the gene bank in E. coli. DNA preparations from these yeast clones yielded no transformants in E. coli but retransformation of yeast was possible. This lead to the isolation of a defective derivative of the rad4 complementing plasmid. The modified plasmid was now capable of transforming E. coli but still interfered significantly with its growth. AU - Siede, W. AU - Eckardt-Schupp, F. C1 - 42581 C2 - 40160 SP - 205-210 TI - DNA repair genes of Saccharomyces cerevisiae: Complementing rad4 and rev2 mutations by plasmids which cannot be propagated in Escherichia coli. JO - Curr. Genet. VL - 11 IS - 3 PY - 1986 SN - 0172-8083 ER - TY - JOUR AB - A diploid yeast thymidylate auxotroph was grown under conditions of thymidylate stress ranging from depletion to excess levels of the nucleotide. High concentrations of thymidylate were mutagenic and recombinagenic whereas starvation for thymine nucleotides was recombinagenic and only slightly mutagenic. These results are discussed in relation to possible mutagenic and recombinagenic mechanisms of nucleotide pool imbalances. AU - Eckardt, F. AU - Kunz, B.A. AU - Haynes, R.H. C1 - 41721 C2 - 38329 SP - 399-402 TI - Variation of mutation and recombination frequencies over a range of thymidylate concentrations in a diploid thymidylate auxotroph. JO - Curr. Genet. VL - 7 IS - 5 PY - 1983 SN - 0172-8083 ER - TY - JOUR AB - Thymine nucleotide starvation is recombinagenic in Saccharomyces cerevisiae and induces formation of the 'nuclear dense body', a structure characteristic of yeast cells in meiosis. Conceivably, thymineless recombination in yeast, presumed to be mitotic, might be meiotic in nature. We have tested this hypothesis and have found that thymineless recombination can be induced in strains incapable of meiotic exchange. AU - Kunz, B.A.* AU - Little, J.G.* AU - Eckardt, F. AU - Haynes, R.H.* C1 - 34131 C2 - 38624 SP - 29-31 TI - Thymineless recombination in Saccharomyces cerevisiae is independent of the ability to undergo meiosis. JO - Curr. Genet. VL - 5 IS - 1 PY - 1982 SN - 0172-8083 ER -