Genes, variant genes and pseudogenes of the human tRNA(Val) gene family. Expression and pre-tRNA maturation in vitro.
Nine different members of the human tRNA(Val) gene family have been cloned and characterized. Only four of the genes code for one of the known tRNA(Val) isoacceptors. The remaining five genes carry mutations, which in two cases even affect the normal three-dimensional tRNA structure. Each of the genes is transcribed by polymerase III in a HeLa cell nuclear extract, but their transcription efficiencies differ by up to an order of magnitude. Conserved sequences immediately flanking the structural genes that could serve as extragenic control elements were not detected. However, short sequences in the 5' flanking region of two genes show striking similarity with sequences upstream from two Drosophila melanogaster tRNA(Val) genes. Each of the human tRNA(Val) genes has multiple, i.e. two to four, transcription initiation sites. In most cases, transcription termination is caused by oligo(T) sequences downstream from the structural genes. However, the signal sequences ATCTT and CTTCTT also serve as effective polymerase III transcription terminators. The precursors derived from the four tRNA(Val) genes coding for known isoacceptors and those derived from two mutant genes are processed first at their 3' and subsequently at their 5' ends to yield mature tRNAs. The precursor derived from a third mutant gene is incompletely maturated at its 3' end, presumably as a consequence of base-pairing between 5' and 3' flanking sequences. Finally, precursors encoded by the genes that carry mutations affecting the tRNA tertiary structure are completely resistant to 5' and 3' processing.
Thomann
H U
HU
Institut für Biochemie, Bayerische Julius-Maximilians-Universität, Würzburg, Federal Republic of Germany.
Schmutzler
C
C
Hüdepohl
U
U
Blow
M
M
Gross
H J
HJ
D001483:Base Sequence; D015870:Gene Expression; D005796:Genes; D006801:Humans; D005810:Multigene Family; D009693:Nucleic Acid Hybridization; D009710:Nucleotide Mapping; D012322:RNA Precursors; D012345:RNA, Transfer, Amino Acid-Specific / Q000032:analysis*; D012366:RNA, Transfer, Val / Q000032:analysis* / Q000235:genetics; D013728:Terminator Regions, Genetic; D014158:Transcription, Genetic
D012322:RNA Precursors; D012345:RNA, Transfer, Amino Acid-Specific; D012366:RNA, Transfer, Val