Toxoplasma gondii ribonuclease P (RNase P) activity was identified and
characterized. The enzyme cleaved the precursor of Escherichia coli t
yrosine transfer RNA at a position identical to the site recognized by
E. coli RNase P. The enzyme has a broad pH optimum from 7 to 9, is mo
re active at 42 degrees C than at 32 degrees C and 37 degrees C, and i
s relatively stable at 37 degrees C and 42 degrees C, but not at 56 de
grees C. Its activity is stimulated significantly as the concentration
of Mg+2 is increased to 100 mM, but some activity remains in the pres
ence of less than 1 mu M free Mg+2; Ca+2 stimulates activity modestly
up to 25 mM. Toxoplasma gondii RNase P differs from host (murine perit
oneal exudate cells) RNase P, as indicated by its elution from cationi
c ion exchange columns at a relatively high salt concentration. It is
effectively inhibited by mature tRNA and inactivated by pretreatment w
ith micrococcal nuclease, thus indicating the presence of an essential
RNA component. Correspondingly, a number of RNA molecules, ranging in
size from about 170 to 490 nucleotides, are found in T. gondii RNase
P containing fractions. This study provides the basis for further char
acterization of the structure and function of T. gondii RNase P and it
extends the range of organisms in which RNase P has been characterize
d.