Reduced dosage of the chromosome axis factor Red1 selectively disrupts the meiotic recombination checkpoint in Saccharomyces cerevisiae
Fig 4
red1ycs4S mutants lack a sustained meiotic DNA damage checkpoint arrest.
(A) Sporulation efficiency of red1ycs4S (H7011), rec8Δ (H5187), and red1ycs4S rec8Δ (H7661) mutants relative to wild type (H7797), n = 200. (B) Western analysis of phosphorylation levels of Hop1-T318 and H3-T11 from wild-type and red1ycs4S cells. Fpr3 was used as loading control. (C) Spindle pole separation in fixed cells measured by anti-tubulin immunofluorescence of wild type (black circle), red1ycs4S (cyan triangle), rec8Δ (pink square), and red1ycs4S rec8Δ (orange diamond), n = 200. Spindle pole separation is an indication of progression out of meiotic prophase. (D) Spindle pole separation in red1ycs4S (H7088), rec8Δ (H7161), and red1ycs4S rec8Δ (H6589) mutants and their control (H7076) in a repair-deficient dmc1Δ rad51Δ background, n = 200. (E) Spindle pole separation of wild type (black closed circle), red1ycs4S (cyan closed triangle), red1-pG162A (H9080, cyan open triangle) and its matched control (H9078, black open circle) strains in a dmc1Δ rad51Δ background to measure checkpoint activity, n = 200. (F) Spindle pole separation of wild type (black closed circle), red1ycs4S (cyan closed triangle), pHOP1-RED1 (H8851, black open circle), and pHOP1-RED1ycs4S (H8852, cyan open triangle) strains in a dmc1Δ rad51Δ background to measure checkpoint activity, n = 200.