Pellegrini Olivier, Nezzar Jamel, Marchfelder Anita, Putzer Harald, Condon Ciarán
CNRS UPR 9073, Institut de Biologie Physico-Chimique, 13 rue Pierre et Marie Curie, 75005 Paris, France.
EMBO J. 2003 Sep 1;22(17):4534-43. doi: 10.1093/emboj/cdg435.
In contrast to Escherichia coli, where the 3' ends of tRNAs are primarily generated by exoribonucleases, maturation of the 3' end of tRNAs is catalysed by an endoribonuclease, known as RNase Z (or 3' tRNase), in many eukaryotic and archaeal systems. RNase Z cleaves tRNA precursors 3' to the discriminator base. Here we show that this activity, previously unsuspected in bacteria, is encoded by the yqjK gene of Bacillus subtilis. Decreased yqjK expression leads to an accumulation of a population of B.subtilis tRNAs in vivo, none of which have a CCA motif encoded in their genes, and YqjK cleaves tRNA precursors with the same specificity as plant RNase Z in vitro. We have thus renamed the gene rnz. A CCA motif downstream of the discriminator base inhibits RNase Z activity in vitro, with most of the inhibition due to the first C residue. Lastly, tRNAs with long 5' extensions are poor substrates for cleavage, suggesting that for some tRNAs, processing of the 5' end by RNase P may have to precede RNase Z cleavage.
与大肠杆菌不同,在大肠杆菌中tRNA的3'末端主要由外切核糖核酸酶产生,而在许多真核生物和古细菌系统中,tRNA 3'末端的成熟是由一种称为RNase Z(或3' tRNase)的内切核糖核酸酶催化的。RNase Z在鉴别碱基的3'端切割tRNA前体。我们在此表明,这种以前在细菌中未被怀疑的活性由枯草芽孢杆菌的yqjK基因编码。yqjK表达降低导致枯草芽孢杆菌体内一群tRNA积累,这些tRNA的基因中均未编码CCA基序,并且YqjK在体外以与植物RNase Z相同的特异性切割tRNA前体。因此,我们将该基因重新命名为rnz。鉴别碱基下游的CCA基序在体外抑制RNase Z活性,大部分抑制作用归因于第一个C残基。最后,具有长5'延伸的tRNA是切割的不良底物,这表明对于某些tRNA,RNase P对5'端的加工可能必须先于RNase Z切割。