A spermidine-dependent endoribonuclease (designated as RNase 65) activity requires both RNA and protein components (Nashimoto et al. (1991) Biochem. Biophs. Res. Comm. 176:1163-1169). In this study, we fractionated RNAs from mouse FM3A cell extracts and showed that an RNA fraction containing two major RNAs and two minor ones restored the micrococcal nuclease-inhibited RNase 65 activity. Partial sequences of these four RNA species were determined by chemical RNA sequencing. A sequence homology search revealed that the two major RNAs were glutamine tRNA lacking its 3' terminus, and that the two minor RNAs were initiator methionine tRNA and glycine tRNA lacking their 3' termini.