Genes coding for valine transfer ribonucleic acid-3b in Drosophila melanogaster.
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Animals↗
Biomedical subjects
G M Tener
Publications and source records attributed to G M Tener.
Isolation and characterization of recombinant DNA plasmids carrying Drosophila tRNA genes.
Recombinant plasmids carrying Drosophila melanogaster tRNA genes were constructed by ligation of HindIII-cleaved Drosophila DNA to HindIII cut pBR322 DNA. 90 clones were isolated that contained genes for one or more of eleven tRNAs. 43 of the plasmids were characterized by a number of methods: restriction nuclease digestion; agarose gel electrophoresis; hybridization with individual, purified, 125I-labelled Drosophila tRNA molecules and in situ hybridization to Drosophila chromosomes. The results show that several different tRNA genes have been isolated which code for single, specific isoacceptors. The DNAs from 8 plasmids each hybridize to single sites on Drosophila polytene chromosomes. In addition, the data show examples of two different plasmids hybridizing to different loci coding for the same tRNA; this means that we have isolated representatives of tRNA genes which map at widely separated points on the Drosophila genome.
Animals↗
The nucleotide sequence of the initiator tRNA from Drosophila melanogaster.
The nucleotide sequence of Drosophila melanogaster methionine tRNAi was determined to be: pA-G-C-A-G-A-G-U-m1G-m2G-C-G-C-A-G-U-G-G-A-A-G-C-G-U-m2G-C-U-G-G-G-C-C-C-A-U-t6A-A-C-C-C-A-G-A-G-m7G-D-m5C-C-C-G-A-G-G-A-U-C-G-m1A-A-A-C-C-U-U-G-C-U-C-U-G-C-U-A-C-C-A(OH). It differs from vertebrate initiator tRNAs in only 6 out of 75 positions.
Animals↗
The nucleotide sequence of lysine tRNA2 from Drosophila.
The nucleotide sequence of Drosophila melanogaster lysine tRNA2 was determined to be: pG-C-C-C-G-G-C-U-A-m2G-C-U-C-A-G-D-C-G-G-D-A-G-A-G-C-A-psi-G-A-G-A-C-U-C-U-U-t6A-A-psi-C-U-C-A-G-G-m7G-D-C-G-U-G-G-G-Xm-U-C-G-m1A-G-C-C-C-C-A-C-G-U-U-G-G-G-C-G-C-C-A(OH). With minor differences in the state of modification of some nucleotides, the sequence is the same as that of lysine tRNA2b from rabbit liver.
Animals↗
Synthesis of 125I-labeled N-3-(4-hydroxyphenyl)propionyl aminoacyl transfer ribonucleic acids.
A method for the isolation and labeling to high specific radioactivity of individual isoaccepting tRNAs is described. After blocking reactive minor bases by acetylation and iodination of the crude tRNA, a single family of isoacceptors was aminoacylated. Individual isoacceptors were separated by chromatography on RPC-5 and then acylated with the 3-(4-hydroxyphenyl)propionyl ester of N-hydroxysuccinimide. The product was purified by chromatography on BD-cellulose and RPC-5. This derivatized tRNA was then iodinated with 125I- and Chloramine-T to give a product containing between 5 X 10(7) and 3 X 10(8) dpm/microgram. The suitability of such labeled tRNAs for hybridization to homologous DNA in solution and cytological preparations of chromosomes is discussed with particular reference to Drosophila melanogaster.
Chromatography, Gel↗
The purification and properties of valine tRNAs of Drosophila melanogaster.
The valine transfer ribonucleic acids of Drosophila melanogaster have been purified by column chromatography on BD-cellulose, Sepharose 6B, and RPC-5. Three major species were analyzed for base composition and coding properties. Valyl-tRNAVal3a binds strongly to ribosomes in the presence of GUA and to a lesser extent with GUU and GUG. Valyl-tRNAVal3b binds strongly in the presence of GUG and very poorly if at all with the other three triplets whereas valvyl-tRNAVal4, which contains inosine, binds strongly in the presence of GUU, GUC, and GUA and weakly with GUG.
Animals↗
Acetylation of chromosome squashes of Drosophila melanogaster decreases the background in autoradiographs from hybridization with [125I]-labeled RNA.
DNA in prepared chromosomes from the larval salivary glands of Drosophila melanogaster was hybridized with [125I]-labeled 5S and tRNA from the same organism. Autoradiography revealed that radioactivity was frequently bound to all regions of the slides, masking labeling of the chromosomes. Acetylation of the preparations before hybridization prevented the formation of this background and revealed the specific chromosomal sites.
Acetylation↗
An analysis of five serine transfer ribonucleic acids from Drosophila.
Crude tRNA from adult Drosophila melanogaster was fractionated on bensoylated-diethylaminoethyl cellulose columns. The eluate was assayed for both amino acid acceptance and cytokinin activity. Most of the cytokinin activity was associated with a peak of serine acceptance. The five major serine tRNAs were purified by chromatography on benzoylated-dietyhlaminoethyl cellulose and reversed phase chromatography-5 columns. The major species, tRNA7-Ser was isolated from this tRNA and was shown to be N-6-(delta-2-isopentenyl)adenosine (i-6A) on the basis of ultraviolet and mass spectral data. The nucleoside somposition of all five serine tRNAs was determined directly and by the 3-H derivative method. They all contain pseudouridine, ribothymidine, 1-methyladenosine, 5-methylcytosine, N-2-dimethylguanosine, 5, 6-hydrouridine, and 3-methylcytosine, while two contain an unidentified nucleoside, and one containes 1-methylguanosine. These techniques also confirmed the presence of i-6A in tRNA7-Ser as well as showing its presence in tRNA6-Ser and tRNA4-Ser. These three tRNA-Ser species exhibit marked changes in elution from reversed phase chromatography-5 columns as a function of temperature and this may be related to their minor base composition. The tRNAs-Ser were bound to ribosomes in response to the following triplets: tRNA2-Ser, AGU, AGC; tRNA4-Ser, UCG; tRNA5-Ser, AGU, AGC; tRNA7-Ser, UCG.
Adenosine↗