Nucleotide sequence of Drosophila melanogaster 5S RNA: Evidence for a general 5S RNA model (original) (raw)
Related papers
Molecular and Cellular Biology, 1987
We constructed deletion-substitution and linker-scanning mutations in the 5'-flanking region of the Drosophila melanogaster 5S RNA gene. In vitro transcription of these templates in Drosophila and HeLa cell extracts revealed the presence of an essential control region (-30 region) located between nucleotides -39 and -26 upstream of the transcription initiation site: deletion of sequences upstream of nucleotide position -39 had no detectable effect on the wild-type level of in vitro transcription, whereas mutations extending between positions -39 and 1 resulted in templates with decreased transcriptional levels; specifically, deletion and linker-scanning mutations in the -34 to -26 region (-30 region) resulted in loss of transcription. The -30 region is essential for transcription and therefore forms part of the Drosophila 5S RNA gene transcription promoter. Compared with the activity of the wild-type gene, mutant 5S DNAs exhibited no impairment in the ability to sequester limiti...
The EMBO Journal, 1982
The organization of the 5S RNA gene cluster of Drosophila melanogaster is different in two Oregon R stocks that have been separated for a number of years. The Oregon R Yale population contains various different arrangements of the cluster. One of these is due to the insertion of a B104 element near one end of the cluster. Other arrngements lack the B104 insertion and have instead a variety of deletions originating in the vicinity of the B104 insertion site and removing from 0 to 6007o of the 5S RNA genes without affecting nearby tRNA genes. In contrast, the Oregon R Heidelberg population has no B104 element in the 5S gene cluster and no heterogeneity in the arrangement of the cluster. We propose that transposable elements inserted at a genomic locus generate heterogeneity in a population at that locus due to excision of the element with and without accompanying deletions of flanking sequences. As a consequence, a fly population would accumulate a large number of deletions scattered throughout the genome in as many loci as contain transposable elements. We show further that D. melanogaster contains a large redundancy of 5S RNA genes since the 600/0 deletion of the cluster shows no visible phenotype when homozygous or when heterozygous against a total deletion of the entire 5S gene cluster.
MGG Molecular & General Genetics, 1976
Based on the comparative analyses of the primary structure of 5 S RNAs from 19 organisms, a secondary structure model of 5S RNA is proposed. 5S RNA has essentially the same structure among all prokaryotic species. The same is true for eukaryotic 5 S RNAs. Prokaryotic and eukaryotic 5 S RNAs are also quite similar to each other, except for a difference in a specific region. By comparing the nucleotide alignment from the juxtaposed 5S RNA secondary structures, a phylogenic tree of nineteen organisms was constructed. The time of divergence between prokaryotes and eukaryotes was estimated to be 2.5 x 109 years ago (minimum estimate: 2.1 x 109).
Position of 5-S RNA among cellular ribonucleic acids
Biochimica et biophysica acta, 1967
A 5-S RNA located on the 5o-S (or 6o-S) ribosomes has been isolated from various uninfected cells. Its relation to other cellular RNA's is not yet established. The present paper reports time-course labeling experiments. Analysis of results by the mathematical procedure of BRITTEN AND MCCARTHY demonstrates that 5-S RNA is not a tRNA precursor. In view of various properties of the 5-S RNA base composition, nature of the 3' and 5' ends, proportion of the total RNA, extractmn patterns, it can be assumed that 5-S RNA is not a degradation product of rRNA and must be considered as a third rRNA.
European Journal of Biochemistry, 1979
The ribonucleoprotein complex between 5-S RNA and its binding protein (5-S RNA . protein complex) of yeast ribosomes was released from 60-S subunits with 25 mM EDTA and the protein component was purified by chromatography on DEAE-cellulose. This protein, designated YL3 (MI = 36000 on dodecylsulfate gels), was relatively insoluble in neutral solutions (pH 4-9) and migrated as one of four acidic 60-S subunit proteins when analyzed by the Kaltschmidt and Wittman two-dimensional gel system. Amino acid analyses indicated lower amounts of lysine and arginine than most ribosomal proteins. Sequence homology was observed in the N terminus of YL3, and two prokaryotic 5-S RNA binding proteins, EL 18 from Escherichia coliand HL 13 from Halobacterium cutirubrum : Ala'-
Evolutionary changes in the higher order structure of the ribosomal 5S RNA
Nucleic Acids Research, 1987
Comparative studies have been undertaken on the higher order structure of ribosomal 5S RNAs from diverse origins. Competitive reassociation studies show that 5S RNA from either a eukaryote or archaebacteriurn will form a stable ribonucleoprotein complex with the yeast ribosomal 5S RNA binding protein (YL3); in contrast, eubacterial RNAs will not compete in a similar fashion. Partial S ribonuclease digestion and ethylnitrosourea reactivity were used to probe the structural differences suggested by the reconstitut ion experiments. The results indicate a more compact higher order structure in eukaryotic 5S RNAs as compared to eubacteria and suggest that the archaebacterial 5S RNA contains features which are common to either group. The potential significance of these results with respect to a generalized model for the tertiary structure of the ribosomal 5S RNA and to the heterogeneity in the protein components of 5S RNA-protein complexes are discussed.
Base sequence complexity of the stable RNA species of Drosophila melanogaster
Biochemistry, 1976
The base sequence complexity of Drosophila transfer RNA (tRNA), 5 s RNA, and 1 8 s + 2 8 s ribosomal RNA was determined by analyzing the kinetics of RNA-DNA hybridization on membrane filters. We find that Drosophila tRNA is made up from about 59 basic nucleotide sequences distinguishable by hybridization, suggesting that many of the D r o s o p h i l a melanogaster is one of the few eukaryotes in which the transfer RNAs for all 20 amino acids have been thoroughly analyzed by reverse phase chromatography (RPC).' White et al. (1973a) have shown that Drosophila tRNA can be resolved on RPC-5 columns (Pearson et al., 197 1) into 99 peaks. The pattern is somewhat more complex than that found in Escherichia coli, human, or mouse, where about 56 distinct components are resolved (Gallo and Pestka, 1970). White et al. (1973b) have evidence which suggests that several chromatographically distinct forms of isoaccepting tRNAs essentially have the same nucleotide sequence and are probably products of the same genes. The conversion of one form to another is believed to be mediated by a t R N A modifying enzyme which modifies a single nucleotide. These authors proposed the term "homogenic" to describe tRNAs presumably transcribed from the same genes which are chromatographically distinct as a result of different degrees of posttranscriptional modification. These findings raise the possibility that other isoaccepting forms of tRNA might also be homogenic. In order to investigate this possibility, the kinetic hybridization technique of Birnstiel et al. (1972) was employed in this study to examine the sequence complexity of 4 s RNA of Drosophila melanogaster. In this method, the time course of hybridization of a
Evolution of 5S rRNA gene families in Drosophila
Chromosome Research
In Drosophila virilis, the three clusters of 5S rRNA genes on chromosome 5 comprise two different gene families (B and C), which differ profoundly in the organization of their spacer sequences. While C-type genes, which are found in two of the clusters, exhibit a true repetitive character, the B-type genes of the third cluster are each embedded in completely different genomic environments. Southern blots of genomic DNA of different D. virilis subspecies, D. hydei and D. melanogaster probed with 5S rRNA gene spacer and coding sequences demonstrate the specificity of C-type sequences for the D. virilis species group. The comparative analysis of flanking sequences of 5S rRNA genes of D. virilis, members of the D. melanogaster species subgroup and of the blowfly Calliphora erythrocephala reveals the existence of conserved sequence motifs both in the 5' upstream and 3' downstream flanking regions. Their possible roles in the control of expression and processing of the 5S rRNA pre...