Comparative study of levels of secondary processing in bulk mRNA from dry and germinating wheat embryos.

There has been no previous study of levels of secondary processing in the template-active RNA of dry seeds and embryos. Such information is needed to evaluate the role of transcription in changing the cell-free translational capacity of bulk RNA during early imbibition of water by dry wheat embryos [J. Biol. Chem. 255, 5969-5970 (1980)]. Although it probably contains a higher proportion of "hidden breaks' than bulk mRNA from imbibing embryos, bulk mRNA in dry wheat embryos is also "capped' by 7-methylguanosine at 5' termini, devoid of unmethylated "caps', methylated internally (N6-methyladenosine) and polyadenylated at 3' termini. In contrast to other developing systems, there is no evidence that there are signal changes in levels of secondary processing in the bulk mRNA populations which support change in cell-free translational capacity of RNA 1-5 h postimbibition of dry wheat embryos. Change in the pattern of protein synthesis 5-24 h postimbibition also takes place without signal changes in levels of secondary processing in template-active RNA. The analytical data are evaluated and discussed in terms of difficulties allied with comparative analyses of poly(A)-rich RNA from dry and imbibing embryos, the former being refractory to, and the latter being most easily analyzed by, labeling in vivo.

Presence of the methylester of 5-carboxymethyl uridine in the wobble position of the anticodon of tRNAIII Arg from brewer's yeast.

The methylester of 5-carboxymethyluridine (mcm5U), its degradation product 5-carboxymethyluridine (cm5U) and the corresponding nucleotide (cm5Up) were isolated from brewer's yeast tRNAIII Arg or from the dodecanucleotide containing the anticodon. Their chromatographic and electrophoretic properties and their UV absorbing spectra were identical to that of the corresponding synthetic compounds. The gas chromatographic behavior and the mass spectrum of mcm5U obtained from tRNAIII Arg and of a synthetic sample were also identical ; the rare occurence of a thermal reciprocal bimolecular methyl-hydrogen transfer in the mass spectrometer ion source was observed. A mild alkaline treatment of tRNAIII Arg leads to the saponification of mcm5U into cm5U (within the tRNA), which can be again esterified in the presence of a yeast homogenate and (methyl-14C) S adenosylmethionine. The radioactivity was found in the mcm5U located in the wobble position of the anticodon of tRNAIII Arg. The presence of this odd nucleotide in that position could possibly restrict the codon-anticodon interaction of tRNAIII Arg.