Figure 3
ZmASCH exhibits a ribonuclease activity. (a) EMSA with four types of nucleic acids. Each of the indicated nucleic acids (3 nM) was incubated with wild-type ZmASCH (300 nM). EMSA was performed on a 15% (w/v) non-denaturating polyacrylamide gel. (b) 32P-labelled 30-mer RNA (3 nM) was incubated with ZmASCH (300 nM) for 30 min at 310 K in the absence or presence of 10 mM MgCl2, MnCl2, NiCl2, CaCl2, ZnCl2 or EDTA. EMSA was performed on a 15% (w/v) non-denaturating polyacrylamide gel. (c,d) The indicated RNA probes (3 nM) were incubated with wild-type ZmASCH protein (0, 150, 300, 500 nM) for 30 min at 310 K in the presence of 10 mM MgCl2. EMSA was performed on a 20% (w/v) non-denaturating polyacrylamide gel. The brackets (a–d) indicate the degradation products.
