Summary
Transfer RNA was extracted from 50–300 mg of adult flies and specifically labeled in vitro. The level of individual isoacceptors was quantitated by efficient annealing to Drosphila tRNA genes carried on recombinant DNA plasmids immobilized on nitrocellulose filters. The level of tRNA Val3b in the tRNA isolated from flies deficient in the major tRNA Val3b loci has been examined. The results show that deletion of the major tRNA Val3b loci resulted in a reduction of approximately 50% in the level of tRNA Val3b but did not produce the Minute phenotype; furthermore the effects of deficiencies at two loci were approximately additive.
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References
Addison WR, Astell CR, Delaney AD, Gillam IC, Hayashi S, Miller RC, Rajput B, Smith M, Taylor DM, Tener GM (1981) The structures of genes hybridizing with tRNA Val4 from Drosophila melanogaster. J. Biol Chem 257:670–673
Anderson WF (1969) The effect of tRNA concentration on the rate of protein synthesis. Proc Natl Acad Sci USA 62:566–573
Anderson WF, Gilbert JM (1969) tRNA-dependent translational control of in vitro hemoglobin synthesis. Biochem Biophys Res Commun 36:456–462
Atwood KC (1968) In: Lindsley DL, Grell EH (eds) Genetic variations of Drosophila melanogaster. Carnegie Inst Washington Publ. 627, p 152
Champe SP, Benzer S (1962) Reversal of mutant phenotypes by 5-fluorouracil: an approach to nucleotide sequences in messenger-RNA. Proc Natl Acad Sci USA 48:532–546
Chernyshev AI, Bashkirov VN, Leibovitch BA, Khesin RB (1980) Increase in the number of histone genes in case of their deficiency in Drosophila melanogaster. Mol Gen Genet 178:663–668
Delaney A, Dunn R, Grigliatti TA, Tener GM, Kaufman TC, Suzuki DT (1976) Quantitation and localization of tRNA genes of Drosophila melanogaster. Fed Proc 35:1676
Dingermann T, Sharp S, Appel B, DeFranco D, Mount S, Heiermann R, Pongs O, Söll D (1981) Transcription of cloned tRNA and 5S RNA in a Drosophila cell free extract. Nucl Acids Res 9:3907–3918
Duncan IW, Kaufman TC (1975) Cytogenetic analysis of chromosome 3 in Drosophila melanogaster: mapping of the proximal protion of the right arm. Genetics 80:733–752
Dunn R, Addison WR, Gillam IC, Tener GM (1978) The purification and properties of valine tRNAs of Drosophila melanogaster. Can J Biochem 56:618–623
Dunn R, Delaney AD, Gillam IC, Hayashi S, Tener GM, Grigliatti TA, Misra V, Spurr MG, Taylor DM, Miller RC, Jr (1979b) Isolation and characterization of recombinant plasmids carrying Drosophila tRNA genes. Gene 7:197–215
Dunn R, Hayashi S, Gillam IC, Delaney AD, Tener GM, Grigliatti TA, Kaufman TC, Suzuki DT (1979a) Genes coding for valine transfer ribonucleic acids-3b in Drosophila melanogaster. J Mol Biol 128:277–287
Elder RT (1978) Genes for a single transfer RNA are present at two chromosomal sites in Drosophila melanogaster. Fed Proc 37:1732
Endow SA (1980) On ribosomal gene compensation in Drosophila. Cell 22:149–155
Garber RL, Gage LP (1979) Transcription of a cloned Bombyx mori tRNA Ala2 gene: nucleotide sequence of the tRNA precursor and its processing in vitro. Cell 18:817–828
Garel JP, Garber RL, Siddiqui MAQ (1977) Transfer RNA in the posterior silk gland of Bombyx mori: polyacrylamide gel mapping of mature transfer RNA, identification and partial structural characterization of major isoacceptor species. Biochemistry 16:3618–3624
Garel JP, Mandel P, Chavancy G, Daillie J (1970) Functional adaptation of tRNAs to fibroin biosynthesis in the silkgland of Bombyx mori. FEBS Lett 7:327–329
Grigliatti TA, White BN, Tener GM, Kaufman TC, Suzuki DT (1974) The localization of transfer RNA Lys5 genes in Drosophila melanogaster. Proc Natl Acad Sci USA 71:3527–3531
Hayashi S, Addison WR, Gillam IC, Grigliatti TA, Tener GM (1981) Hybridization of tRNAs of Drosophila melanogaster to the region of the 5S RNA genes of the polytene chromosomes. Chromosoma 82:385–397
Hayashi S, Gillam IC, Delaney AD, Dunn R, Tener GM, Grigliatti TA, Suzuki DT (1980) Hybridization of tRNAs of Drosophila melanogaster to polytene chromosomes. Chromosoma 76:65–84
Hershey ND, Davidson N (1980) Two Drosophila melanogaster tRNAGly genes are contained in a direct duplication at chromosomal locus 56F. Nucl Acids Res 8:4899–4910
Hosbach HA, Silberklang M, McCarthy BJ (1980) Evolution of a D. melanogaster glutamate tRNA gene cluster. Cell 21:169–178
Hovemann B, Sharp S, Yamada H, Söll D (1980) Analysis of a Drosophila tRNA gene cluster. Cell 19:889–895
Jacobson KB (1978) Mechanism of suppression in Drosophila VII. Correlation between disappearance of an isoacceptor of tyrosine tRNA and activation of the vermilion locus. Nucl Acids Res 5:2391–2404
Kubli E, Schmidt T (1978) The localization of tRNA Glu4 genes from Drosophila melanogaster by in situ hybridization. Nucl Acids Res 5:1465–1478
Lindsley D, Grell EH (1968) Genetic variations of Drosophila melanogaster. Carnegie Inst Wash Publ 627
Lindsley DL, Sandler L, Baker BS, Carpenter ATC, Denell RE, Hall JC, Jacobs PA, Miklos GLG, Davis BK, Gethmann RW, Hardy RW, Hessler A, Miller SM, Nozawa H, Parry DM, Gould-Somero M (1972) Segmental aneuploidy and the genetic gross structure of the Drosophila genome. Genetics 71:157–184
Miller RC Jr, Young ET II, Epstein RH, Krisch HM, Mattson T, Bolle A (1981) Regulation of the synthesis of the T4 DNA polymerase (Gene 43). Virology 110:98–112
Procunier JD, Tartof KD (1975) Genetic analysis of the 5S RNA genes in Drosophila melanogaster. Genetics 81:515–523
Ritossa FM, Atwood KC, Spiegelman S (1966a) On the redundancy of DNA complementary to amino acid transfer RNA and its absence from the nucleolar organizer region of Drosophila melanogaster. Genetics 54:663–676
Ritossa FM, Atwood KC, Spiegelman S (1966b) A molecular explanation of the bobbed mutants of Drosophila as partial deficiencies of “ribosomal” DNA. Genetics 54:819–834
Robinson RR, Davidson N (1981) Analysis of a Drosophila melanogaster tRNA gene cluster: two tRNALeu genes contain intervening sequences. Cell 23:251–259
Roe BA (1975) Studies on human tRNA. I. The rapid, large scale isolation and partial fractionation of placenta and liver tRNA. Nucl Acids Res 2:21–42
Schmidt T, Egg AH, Kubli E (1978) The localization of tRNA Asp2 genes from Drosophila melanogaster by in situ hybridization. Mol Gen Genet 164:249–254
Schmidt T, Kubli E (1980) The localization of tRNA Asn5 , tRNAHis, and tRNAAla genes from Drosophila melanogaster by in situ hybridization to polytene salivary gland chromosomes. Chromosoma 80:277–287
Twardzik DR, Grell EH, Jacobson KB (1971) Mechanism of suppression in Drosophila: a change in tyrosine transfer RNA. J Mol Biol 57:231–245
White BN, Tener GM, Holden J, Suzuki DT (1973) Analysis of tRNAs during the development of Drosophila. Dev Biol 33:185–195
Yen PH, Davidson N (1980) The gross anatomy of a tRNA gene cluster at region 42A of the D. melanogaster chromosome. Cell 22:137–148
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Communicated by M.M. Green
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Larsen, T.M., Miller, R.C., Spiegelman, G.B. et al. RNA-DNA hybridization analyses of tRNA Val3b in Drosophila melanogaster . Mol Gen Genet 185, 390–396 (1982). https://doi.org/10.1007/BF00334129
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DOI: https://doi.org/10.1007/BF00334129