Journal: Molecular Biology of the Cell
Article Title: Mps1 promotes poleward chromosome movements in meiotic prometaphase
doi: 10.1091/mbc.E20-08-0525-T
Figure Lengend Snippet: Mps1 promotes microtubule turnover in meiotic prometaphase. (A) In wild-type cells the shortening kinetochores of actively biorienting chromosomes are predicted to cause a high microtubule turnover. MPS1 mutants exhibit a locked-in-place phenotype that might represent a defect in the depolymerization of kinetochore microtubules. (B) Cells that were unable to form bipolar attachments ( spo11 ), and thus in a prolonged prometaphase-like state, were used to measure microtubule turnover. Half spindles of meiotic cells were pulsed with 405 nm light to photoconvert mEos2-Tub1 (from green to red). Images were acquired every 15 s, and the intensity of the red signal was measured (see Materials and Methods ). Scale bar: 2 μm. (C) Microtubule turnover on metaphase spindles was measured in a diploid strain undergoing either meiosis or mitosis. (D) Microtubule turnover was measured in cells expressing STU2-AID* in the presence or absence of auxin and CuSO 4 (copper was used to induce expression of the P CUP1 -AFB2 F-box protein construct). (E) Microtubule turnover was measured on meiotic metaphase spindles of wild-type or mps1-as1 cells in the presence of the Mps1-as1 inhibitor 1-NMPP1. (F) Microtubule turnover was measured on meiotic metaphase and prometaphase spindles of wild-type cells. (G) Microtubule turnover was measured on prometaphase spindles ( spo11 ) in cells with or without the inactivation of Mps1 by 1-NMPP1. (H) Microtubule turnover was measured on prometaphase spindles ( spo11 ) in wild-type or mps1-R170S cells. All experiments show the averages and SEM of three or more biological replicates with three or more cells per replicate (see ).
Article Snippet: Where indicated, auxin (2 mM; Sigma Aldrich I5148-10G), CuSO 4 (200 μM; Sigma Aldrich 451657-10G), or 1-NMPP1 (5 μM; Calbiochem; 5 mM stock in dimethyl sulfoxide) were added to the medium at the time of prophase exit.
Techniques: Expressing, Construct