scholarly journals A new method to efficiently induce a site-specific double-strand break in the fission yeast Schizosaccharomyces pombe

Yeast ◽  
2012 ◽  
Vol 29 (7) ◽  
pp. 275-291 ◽  
Author(s):  
Sham Sunder ◽  
Nikole T. Greeson-Lott ◽  
Kurt W. Runge ◽  
Steven L. Sanders
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Strand Break ◽  
Double Strand Break ◽  
New Method ◽  
Site Specific ◽  
A Site

scholarly journals Pathway utilization in response to a site-specific DNA double-strand break in fission yeast

2003 ◽  
Vol 22 (6) ◽  
pp. 1419-1430 ◽  
Author(s):  
J. Prudden
Keyword(s):  
Fission Yeast ◽  
Strand Break ◽  
Double Strand Break ◽  
Site Specific ◽  
Dna Double Strand Break ◽  
A Site

Enzymatic probing and structural mapping of a site-specific complex radiation-induced DNA double-strand break

2003 ◽  
Vol 57 (2) ◽  
pp. S346 ◽  
Author(s):  
K Datta ◽  
M Dizdaroglu ◽  
P Jaruga ◽  
R.D Neumann ◽  
T.A Winters
Keyword(s):  
Strand Break ◽  
Double Strand Break ◽  
Structural Mapping ◽  
Site Specific ◽  
Dna Double Strand Break ◽  
Specific Complex ◽  
Radiation Induced ◽  
A Site

scholarly journals Molecular Analysis of Base Damage Clustering Associated with a Site-Specific Radiation-Induced DNA Double-Strand Break

2006 ◽  
Vol 166 (5) ◽  
pp. 767-781 ◽  
Author(s):  
Kamal Datta ◽  
Pawel Jaruga ◽  
Miral Dizdaroglu ◽  
Ronald D. Neumann ◽  
Thomas A. Winters
Keyword(s):  
Molecular Analysis ◽  
Strand Break ◽  
Double Strand Break ◽  
Base Damage ◽  
Site Specific ◽  
Dna Double Strand Break ◽  
Radiation Induced ◽  
A Site

Site-specific mutagenesis of cdc2+, a cell cycle control gene of the fission yeast Schizosaccharomyces pombe

1986 ◽  
Vol 6 (10) ◽  
pp. 3523-3530
Author(s):  
R Booher ◽  
D Beach
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Cell Cycle Control ◽  
Temperature Sensitive ◽  
Protein Kinase Activity ◽  
Atp Binding Site ◽  
Leu2 Gene ◽  
Cdc2 Gene ◽  
A Site

The cdc2+ gene of Schizosaccharomyces pombe is homologous to the CDC28 gene of Saccharomyces cerevisiae. Both genes share limited homology with vertebrate protein kinases and have protein kinase activity. cdc2+ has been subjected to mutagenesis in vitro. A null allele of the gene, constructed by insertion of the S. cerevisiae LEU2 gene into a site within the gene, has a phenotype similar to that of many temperature-sensitive alleles of cdc2. Mutations within the predicted ATP-binding site and in a region which may be a site of phosphorylation result in loss of cdc2+ activity. A single substitution of Gly-146 to Asp-146 has been identified in cdc2-1w, a dominant activated allele of the gene. The four introns within the cdc2+ gene have been deleted. The resulting gene not only functions in fission yeast but also rescues cdc28(Ts) strains of S. cerevisiae, a property which is not shared by the genomic cdc2+ gene.

scholarly journals Lethality induced by a single site-specific double-strand break in a dispensable yeast plasmid.

1993 ◽  
Vol 90 (12) ◽  
pp. 5613-5617 ◽  
Author(s):  
C. B. Bennett ◽  
A. L. Lewis ◽  
K. K. Baldwin ◽  
M. A. Resnick
Keyword(s):  
Strand Break ◽  
Double Strand Break ◽  
Single Site ◽  
Yeast Plasmid ◽  
Site Specific

scholarly journals Molecular Characterization of the Role of the Schizosaccharomyces pombe nip1+/ctp1+ Gene in DNA Double-Strand Break Repair in Association with the Mre11-Rad50-Nbs1 Complex

2008 ◽  
Vol 28 (11) ◽  
pp. 3639-3651 ◽  
Author(s):  
Yufuko Akamatsu ◽  
Yasuto Murayama ◽  
Takatomi Yamada ◽  
Tomofumi Nakazaki ◽  
Yasuhiro Tsutsui ◽  
...  
Keyword(s):  
Dna Repair ◽  
Schizosaccharomyces Pombe ◽  
Sequence Similarity ◽  
Strand Break ◽  
Double Strand Break ◽  
Double Strand Break Repair ◽  
Dna Double Strand Breaks ◽  
Epistasis Analysis ◽  
Dna Double Strand Break ◽  
Break Repair

ABSTRACT The Schizosaccharomyces pombe nip1 +/ctp1 + gene was previously identified as an slr (synthetically lethal with rad2) mutant. Epistasis analysis indicated that Nip1/Ctp1 functions in Rhp51-dependent recombinational repair, together with the Rad32 (spMre11)-Rad50-Nbs1 complex, which plays important roles in the early steps of DNA double-strand break repair. Nip1/Ctp1 was phosphorylated in asynchronous, exponentially growing cells and further phosphorylated in response to bleomycin treatment. Overproduction of Nip1/Ctp1 suppressed the DNA repair defect of an nbs1-s10 mutant, which carries a mutation in the FHA phosphopeptide-binding domain of Nbs1, but not of an nbs1 null mutant. Meiotic DNA double-strand breaks accumulated in the nip1/ctp1 mutant. The DNA repair phenotypes and epistasis relationships of nip1/ctp1 are very similar to those of the Saccharomyces cerevisiae sae2/com1 mutant, suggesting that Nip1/Ctp1 is a functional homologue of Sae2/Com1, although the sequence similarity between the proteins is limited to the C-terminal region containing the RHR motif. We found that the RxxL and CxxC motifs are conserved in Schizosaccharomyces species and in vertebrate CtIP, originally identified as a cofactor of the transcriptional corepressor CtBP. However, these two motifs are not found in other fungi, including Saccharomyces and Aspergillus species. We propose that Nip1/Ctp1 is a functional counterpart of Sae2/Com1 and CtIP.

scholarly journals Genetic and physical analysis of double-strand break repair and recombination in Saccharomyces cerevisiae.

Genetics ◽  
1989 ◽  
Vol 122 (3) ◽  
pp. 519-534 ◽  
Author(s):  
N Rudin ◽  
E Sugarman ◽  
J E Haber
Keyword(s):  
Strand Break ◽  
Double Strand Break ◽  
The Other ◽  
Crossing Over ◽  
Physical Analysis ◽  
Wild Type ◽  
Inverted Orientation ◽  
Mating Type Switching ◽  
A Site ◽  
Kinetics Of

Abstract We have investigated HO endonuclease-induced double-strand break (DSB) recombination and repair in a LACZ duplication plasmid in yeast. A 117-bp MATa fragment, embedded in one copy of LACZ, served as a site for initiation of a DSB when HO endonuclease was expressed. The DSB could be repaired using wild-type sequences located on a second, promoterless, copy of LACZ on the same plasmid. In contrast to normal mating-type switching, crossing-over associated with gene conversion occurred at least 50% of the time. The proportion of conversion events accompanied by exchange was greater when the two copies of LACZ were in direct orientation (80%), than when inverted (50%). In addition, the fraction of plasmids lost was significantly greater in the inverted orientation. The kinetics of appearance of intermediates and final products were also monitored. The repair of the DSB is slow, requiring at least an hour from the detection of the HO-cut fragments to completion of repair. Surprisingly, the appearance of the two reciprocal products of crossing over did not occur with the same kinetics. For example, when the two LACZ sequences were in the direct orientation, the HO-induced formation of a large circular deletion product was not accompanied by the appearance of a small circular reciprocal product. We suggest that these differences may reflect two kinetically separable processes, one involving only one cut end and the other resulting from the concerted participation of both ends of the DSB.

scholarly journals Yeast ATM and ATR use different mechanisms to spread histone H2A phosphorylation around a DNA double-strand break

2019 ◽  
Author(s):  
Kevin Li ◽  
Gabriel Bronk ◽  
Jane Kondev ◽  
James E. Haber
Keyword(s):  
Strand Break ◽  
Double Strand Break ◽  
Three Dimensions ◽  
One Dimension ◽  
Histone H2a ◽  
Directed Motion ◽  
Checkpoint Kinases ◽  
Dna Double Strand Break ◽  
Chromosome Iii ◽  
A Site

AbstractOne of the hallmarks of DNA damage is the rapid spreading of phosphorylated histone H2A (γ-H2AX) around a DNA double-strand break (DSB). In the budding yeast S. cerevisiae, nearly all H2A isoforms can be phosphorylated, either by Mec1ATR or Tel1ATM checkpoint kinases. We induced a site-specific DSB with HO endonuclease at the MAT locus on chromosome III and monitored the formation of γ-H2AX by ChIP-qPCR in order to uncover the mechanisms by which Mec1ATR and Tel1ATM propagate histone modifications across chromatin. With either kinase, γ-H2AX spreads as far as ∼50 kb on both sides of the lesion within 1 h; but the kinetics and distribution of modification around the DSB are significantly different. The total accumulation of phosphorylation is reduced by about half when either of the two H2A genes is mutated to the nonphosphorylatable S129A allele. Mec1 activity is limited by the abundance of its ATRIP partner, Ddc2. Moreover, Mec1 is more efficient than Tel1 at phosphorylating chromatin in trans – at distant undamaged sites that are brought into physical proximity to the DSB. We compared experimental data to mathematical models of spreading mechanisms to determine whether the kinases search for target nucleosomes by primarily moving in three dimensions through the nucleoplasm or in one dimension along the chromatin. Bayesian model selection indicates that Mec1 primarily uses a 3D diffusive mechanism, whereas Tel1 undergoes directed motion along the chromatin.

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