Fig. 3. bgs4⁺overexpression produces defects in cell integrity. (A) bgs4⁺ overexpression causes cell growth arrest, which is delayed in the presence of sorbitol. Log-phase wild-type and bgs4 p3X-bgs4⁺ cells were grown and monitored after 18 hours of growth in the absence of thiamine as in Fig. 2B. (B) Rounded and elongated morphology of bgs4⁺-overexpressing cells. Log-phase bgs4 p3X-bgs4⁺ cells were grown on EMM liquid medium at 32°C and visualized at the indicated times as in Fig. 1C. (C) bgs4⁺ overexpression induces actin delocalization in rounded cells. bgs4 p3X-bgs4⁺ cells grown on EMM liquid medium at 32°C for 24 hours were stained for actin visualization as in Fig. 2E. (D) bgs4⁺ overexpression produces a gradual reduction in GS activity. Log-phase bgs4 p3X-bgs4⁺ cells grown on EMM as in A were collected at the indicated times and assayed for GS activity as in Fig. 1B. Bar, 10 µm.