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. 1999 Feb;119(2):663–670. doi: 10.1104/pp.119.2.663

Figure 4.

Figure 4

Gel-retardation experiments showing the effects of the preincubation of plastid extracts with either ATP, ADP, phosphocreatine and phosphocreatine kinase, and/or an inhibitor of adenylate kinase. A, Gel-shift assays were done using radiolabeled GT30 DNA fragments and plastid extracts, which were preincubated with either 5.0 mm ATP or 1.0 mm ADP in the presence of 5.0 mm phosphocreatine (PC) and 2 units of phosphocreatine kinase (PC Kin) (lanes 3 and 5, respectively). Binding reactions using plastid extracts preincubated with 5.0 mm ATP or 1.0 mm ADP alone are shown in lanes 2 and 4, respectively. Plastid extracts were obtained from 4.5-d-old, dark-grown barley plants. The major complexes observed using the extracts without any additions (control reaction, lane 1) are labeled I, II, and III. Migration of the free probe (FP) is indicated. B, Gel-retardation experiments using radiolabeled LRP136 DNA probe testing the effect of preincubation of plastid extracts with ADP in the presence of the adenylate kinase inhibitor Ap5A. Plastid extracts were preincubated, before binding reactions, with 1.0 mm Ap5A in the presence (1.0 mm, lane 3) or absence (lane 2) of ADP. The major complexes observed in plastid extracts without additions are labeled A, B, C, D, and E (Control, lane 1). Migration of the free probe (FP) is indicated.