Distribution of both lengths and 5' terminal nucleotides of mammalian pre-tRNA 3' trailers reflects properties of 3' processing endoribonuclease

Abstract

Mammalian tRNA 3' processing endoribonuclease (3' tRNase) removes 3' extra nucleotides after the discriminator from tRNA precursors. Here I examined how the length of a 3' trailer and the nucleotides on each side of the cleavage site affected 3' processing efficiency. I performed in vitro 3' processing reactions of pre-tRNA(Arg)s with various 3' trailers or various discriminator nucleotides using 3' tRNase purified from mouse FM3A cells or pig liver. On the whole, the efficiency of pre-tRNA(Arg) 3' processing by mammalian 3' tRNase decreased as the 3' trailer became longer, except in the case of a 3' trailer composed of CC, CCA or CCA plus 1 or 2 nucleotides, which was not able to be removed at all. The distribution of 3' trailer lengths deduced from mammalian nuclear tRNA genomic sequences reflects this property of 3' tRNase. The cleavage efficiency of pre-tRNA(Arg)s varied depending on the 5' end nucleotide of a 3' trailer in the order G ≃ A > U > C. This effect of the 5' end nucleotide was independent of the discriminator nucleotides. The distribution of the 5' end nucleotides of mammalian pre-tRNA 3' trailers reflects this differential 3' processing efficiency.