Fig 1. The deficits in Lam2, Gtr1 and Gtr2 cause growth defects.

(A) Δlam2 cells, Δgtr1 cells and Δgtr2 cells showed growth defects. Prototrophic wild-type cells (wt, KP5080), Δlam2 cells (KP6578), Δgtr1 cells (KP6573) and Δgtr2 cells (KP6571) were cultured to mid-log phase, and adjusted to 0.3 in OD660. The cells were serially diluted by 10 fold and spotted onto the indicated plates, and were then incubated at 27°C on EMM plates for 4 days and on YES or YPD plates for 3 days. (B) Growth defects of Δlam2 cells, Δgtr2 cells and Δgtr1 cells were rescued by overexpression of the respective genes. The indicated leucine auxotrophic Δlam2 cells (KP6607), Δgtr2 cells (KP6608) and Δgtr1 cells (KP6688) transformed with the control vector (pKB1037) or the plasmid containing the lam2⁺ (pKB8727), gtr2⁺ (pKB8746) or gtr1⁺ (pKB9042) gene were dropped and incubated as described in Fig 1A. (C) tetO₇-lam2 cells and tetO₇-gtr2 cells phenocopied Δlam2 cells and Δgtr2 cells. The Δgtr2 cells (KP6571), tetO₇-gtr2 cells (KP6645), Δlam2 cells (KP6578) and tetO₇-lam2 cells (KP6650) were spotted onto the indicated plates with or without 2.5 μg/ml ahTet, and then incubated as described in Fig 1A. (D) Growth defects of tetO₇-lam2 cells and tetO₇-gtr2 cells were rescued by overexpression of the respective genes. The tetO₇-gtr2 cells (KP6641) transformed with the control vector (pKB1037) or the plasmid containing the gtr2⁺ gene (pKB8746), or the tetO₇-lam2 cells (KP6648) transformed with the control vector (pKB1037) or the plasmid containing the lam2⁺ gene (pKB8727) were spotted and incubated as described in Fig 1A.