Results: 4

1.
Fig. 2.

Fig. 2. From: Regulation of fragile sites expression in budding yeast by MEC1, RRM3 and hydroxyurea.

Effects of rrm3Δ on Sml1 levels and bulk genome duplication in mec1-4ts. (A) MYC–Sml1 levels in the indicated strains following an α-factor arrest and release. The signal corresponding to the MYC–Sml1 is quantified and normalized to that of the tubulin band. Normalized signals are then expressed relative to the t=0 sample, which was set to 1 for each strain. ‘UnT’, untagged control strain. (B) FACS profiles of the samples in A. (C) Status of ChrIII in the indicated strains. Two independently derived rrm3Δ mec1-4ts tel1Δ strains were analyzed.

Nadia Hashash, et al. J Cell Sci. 2011 January 15;124(2):181-185.
2.
Fig. 3.

Fig. 3. From: Regulation of fragile sites expression in budding yeast by MEC1, RRM3 and hydroxyurea.

Effects of HU on RSZ expression in mec1-4ts. α-factor-arrested mec1-4ts cells were released into YPD containing varying concentrations of HU at 30°C (A) or at 23°C (B). Samples were collected at various time points and assessed for: (i) the status of ChrIII; (ii) the number of colony forming units (CFUs); (iii) FACS profiles; and (iv) fraction of cells showing small-budded (open circle) and large-budded (thick line) morphology. Filled circles are CFU data from ii. Asterisks denote the time at which 50% of the culture becomes committed to inviability (T50).

Nadia Hashash, et al. J Cell Sci. 2011 January 15;124(2):181-185.
3.
Fig. 1.

Fig. 1. From: Regulation of fragile sites expression in budding yeast by MEC1, RRM3 and hydroxyurea.

rrm3Δ suppression of mec1-4ts chromosome breakage and temperature sensitivity. (A) ChrIII species revealed by PFGE followed by indirect labelling of one chromosome end (probe: CHA1): full-length linear chromosomes (FL), nonlinear forms that remain in the wells of the gel (Well), and linear chromosome fragments extending from the labelled end (CF). Break positions along the chromosome are deduced from the lengths of CF species. (B) The six RSZs in ChrIII are indicated in roman numerals (I–VI). Numbers below represent the approximate mid-point of each RSZ. Open circle, active ARS. (C,D) Status of ChrIII in the indicated strains. Each lane represents independently derived strains of the specific genotype. Roman numerals correspond to the six RSZs in ChrIII (B). Empty and RRM3 refer to the presence of an ARS-CEN or the same plasmid carrying RRM3. (E) rrm3Δ suppression of mec1-4ts temperature sensitivity.

Nadia Hashash, et al. J Cell Sci. 2011 January 15;124(2):181-185.
4.
Fig. 4.

Fig. 4. From: Regulation of fragile sites expression in budding yeast by MEC1, RRM3 and hydroxyurea.

HU induction of RSZ expression in various checkpoint mutants. (A) α-factor-arrested mec1Δ sml1Δ cells were released into YPD medium containing varying concentrations of HU at 30°C. Samples were collected at the indicated times and subjected to different analyses. (i) Status of ChrIII. (ii) Direct comparison between the distribution of HU induced DSBs in mec1Δ sml1Δ cells and that induced by thermal inactivation of Mec1. Roman numerals correspond to the six RSZs in ChrIII (Fig. 1B). (iii) The number of CFUs. Dotted lines denote the time at which 50% of the culture becomes commitment to inviability (T50). (iv) FACS profiles. (B) Effects of HU on the indicated checkpoint mutants. (C) A model for dNTP regulation of RSZ expression. The extent of dNTP depletion regulates RSZ expression in mec1 or rad53K277A mutants. The overall dNTP levels in each strain are determined by the status of MEC1, SML1 and the concentrations of HU. When dNTP concentrations fall below the viable threshold (asterisks), mec1 or rad53 cells lose viability. If the extent of dNTP depletion is modest (the area marked by the grey box), RSZ expression is observed (grey circles). When the dNTP concentration falls below this level, the cells become prematurely committed to death before RSZ expression (black circles).

Nadia Hashash, et al. J Cell Sci. 2011 January 15;124(2):181-185.

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