FIGURE 4.

Phosphorylation of TMK1 and TMK3 in response to injury and eATP. (A) The WT strain was injured and mycelial samples collected at the indicated times. Mycelium from an undamaged colony was included as control (C). Proteins were extracted, separated by SDS-PAGE, and used for immunoblotting. Blots were probed with anti-Tmk1 (anti-p42/p44) and Tmk1-P (anti-Phospho-p42/p44) antibodies (left panel) or anti-Tmk3 (antip38) and Tmk3-P (anti-Phospho-p38) antibodies (right panel). Note that the anti-Tmk1 antibodies also recognize Tmk2 (p44), as previously shown (Mendoza-Mendoza et al., 2003). (B) The WT strain was ATP induced (0.1 mM) or treated with EGTA (15 mM) and mycelial samples collected at the indicated times. Proteins were extracted, separated by SDS-PAGE, and used for immunoblotting. Blots were probed as in (A). Arrows indicate the bands corresponding to Tmk1 or Tmk3. The Δtmk1 and Δtmk3 mutants were included as controls. The experiments were repeated three times with similar results.