Reduced aminoacylation of asparagine-transfer RNA early in the developmental cycle of Dictyostelium discoideum: modification pattern and possible significance of the uncharged isoacceptor tRNAAsn3.

Changes in the patterns of isoacceptors of tRNAAsn and alterations in modification of the guanine residue 34, the first position of the anticodon of tRNAAsn, have been observed in eukaryotes during differentiation. We use Dictyostelium discoideum as a model system to elucidate the possible involvement of tRNAAsn in developmental processes. Vegetative amoebae were induced to undergo developmental transition by nutrient starvation. Since amino acid starvation alone is a specific stimulus initiating development and unacylated tRNAs might be involved in control mechanisms of protein synthesis, the level of aminoacylation of tRNAAsn isoacceptors has been investigated. As early as two minutes after the onset of development, the aminoacylation of tRNAAsn specifically was reduced to about 30%, whereas at the same time 10 other tRNA species were found to be charged normally, i.e. to 70-100%. One of the two major isoacceptors, tRNAAsn3, was completely deacylated, whereas the other one, tRNAAsn2, accounted for the residual aminoacylation. Analyses of the modified nucleosides of highly purified tRNAAsn2 and tRNAAsn3 are respectively, show that both isoacceptors are identical in their modification patterns except for the modification at the first position of the anticodon; tRNAAsn2 comprises queuine (Q), 7-[(4,5-cis-dihydroxy-2-cyclopenten-1-ylamino)methyl]-7-deazaguanine, whereas tRNAAsn3 contains guanine.