Figure 6.
Figure 6. PB integrity is required for the normal responses and shmoo localization of MFA2 mRNA granules to αF treatment and for efficient mating. (A and B) MFA2 mRNA was expressed in WT cells and in two strains defective in PB formation—pat1Δ dhh1Δ (ySA25; A) and edc3Δ lsm4ΔC (ySA25; B). The cells were treated with αF for 2 h, and the number of MFA2 mRNA granules were counted (n = 200–350 cells). Histograms are plotted as in Fig. 1 B. (C) Granules are preferentially localized in the shmoo-proximal portion of WT cells but not in mutant cells. WT and double deletion mutant strains, described in A and B, were treated with αF for 2, 4, or 6 h. Each type of cell was divided into two equal portions, one containing a shmoo (“shmoo-proximal”) and a “shmoo-distal” (cell body) one. MFA2 mRNA granules were counted and classified into high- and low-intensity granules. We note that the difference in the intensity between the two types of granules is ∼10-fold (see Table 1), and the distinction between them was quite apparent. The ratio between the total number of granules in shmoo-proximal region and in shmoo-distal region is indicated. The ratio of distribution for high (top)- and low (bottom)-intensity granules was calculated for the indicated strains. No high-intensity granules were observed in the double deletion mutant strains. Error bars represent the standard deviation from three independent experiments. n = 100–125 cells in each sample. (D) The studied mutant strains are sterile. The mating assay was determined with the indicated mutant strains (yMC376; yMC524) or isogenic WT (yMC375; all are MATa), as was reported previously (Michaelis and Herskowitz, 1988). The tester strain was WT (MATα; yMC134; see Table 4). After 3 h of mating on rich nonselective plate (YPD), the cells were replica plated onto appropriate selectable plate and incubated for 3 d before the photo was taken. (E) edc3Δ (yMC429) cells are only partially sterile relatively to the edc3Δ lsm4ΔC strain. Semiquantitative mating assay was performed as in D except that 10-fold dilutions were spotted, as indicated.

PB integrity is required for the normal responses and shmoo localization of MFA2 mRNA granules to αF treatment and for efficient mating. (A and B) MFA2 mRNA was expressed in WT cells and in two strains defective in PB formation—pat1Δ dhh1Δ (ySA25; A) and edc3Δ lsm4ΔC (ySA25; B). The cells were treated with αF for 2 h, and the number of MFA2 mRNA granules were counted (n = 200–350 cells). Histograms are plotted as in Fig. 1 B. (C) Granules are preferentially localized in the shmoo-proximal portion of WT cells but not in mutant cells. WT and double deletion mutant strains, described in A and B, were treated with αF for 2, 4, or 6 h. Each type of cell was divided into two equal portions, one containing a shmoo (“shmoo-proximal”) and a “shmoo-distal” (cell body) one. MFA2 mRNA granules were counted and classified into high- and low-intensity granules. We note that the difference in the intensity between the two types of granules is ∼10-fold (see Table 1), and the distinction between them was quite apparent. The ratio between the total number of granules in shmoo-proximal region and in shmoo-distal region is indicated. The ratio of distribution for high (top)- and low (bottom)-intensity granules was calculated for the indicated strains. No high-intensity granules were observed in the double deletion mutant strains. Error bars represent the standard deviation from three independent experiments. n = 100–125 cells in each sample. (D) The studied mutant strains are sterile. The mating assay was determined with the indicated mutant strains (yMC376; yMC524) or isogenic WT (yMC375; all are MATa), as was reported previously (Michaelis and Herskowitz, 1988). The tester strain was WT (MATα; yMC134; see Table 4). After 3 h of mating on rich nonselective plate (YPD), the cells were replica plated onto appropriate selectable plate and incubated for 3 d before the photo was taken. (E) edc3Δ (yMC429) cells are only partially sterile relatively to the edc3Δ lsm4ΔC strain. Semiquantitative mating assay was performed as in D except that 10-fold dilutions were spotted, as indicated.

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