[3] Use of in Vitro32P labeling in the sequence analysis of nonradioactive tRNAs

Silberklang, Melvin; Gillum, Amanda M.; RajBhandary, Uttam L.

doi:10.1016/0076-6879(79)59072-7

Cited by 442 publications

(230 citation statements)

References 55 publications

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“…The purified tRNAGIn was sequenced by the post-labeling procedures described by Silberklang et al ( 11 2), and sequenced by partial digestion with snake venom phosphodiesterase or nuclease P1, followed by two-dimensional homochromatography (I11). Overlapping of the fragments was achieved by partial digestion of 5'-32P-labeled intact tRNA or large tRNA fragments with nuclease P1, or by sequencing gel (13).…”

Section: Methodsmentioning

confidence: 99%

“…This finding was consistent with the HPLC estimate of 1.5 moles of m5C in the molecule. The assignment of mlA to position 57 was based on its characteristic reverse mobility shift on two-dimensional homochromatography and the normalization of its behavior after conversion to m6A (11). Finally, the HPLC analysis indicated that there may be 5 moles of * in the molecule.…”

Section: Methodsmentioning

confidence: 99%

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The nucleotide sequence of a major glutamine tRNA from rat liver

et al. 1983

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

The nucleotide sequence of a major glutamine tRNA from rat liver

et al. 1983

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“…Sequencing of the oligonucleotides and 5'-end labelling of tRNA was performed according to [ 8].3'-End labelling was done as in [9]. The analysis of 2 large fragments isolated from a partial pancreatic digest provided additional results.…”

Section: Methodsmentioning

confidence: 99%

The nucleotide sequence of sheep liver histidine‐tRNA (anticodon Q‐U‐G)

Boisnard

Pétrissant

1981

FEBS Letters

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“…To label the 5' end of MS2 RNA, the latter was treated with alkaline phosphatase [41]. After deproteinisation 10 -20 pmol of MS2 RNA was mixed with 10 pmol of [y-32P]ATP and 1-5 units of T4 polynucleotide kinase.…”

Section: -S Subunits Of E Coli Ribosomes Form Complexes Withmentioning

confidence: 99%

Proteins of the 30‐S Subunit of Escherichia coli Ribosomes which Interact Directly with Natural mRNA

Broude

Kussova

Mecvedeva

et al. 1983

European Journal of Biochemistry

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Ultraviolet irradiation (254 nm) of the complexes of MS2 phage RNA (mRNA) and the 30-S subunits of E.sclzeviclzia coli ribosomes prepared at 0 'C and 37 'C in the presence and absence of initiation factor 3 (IF-3) causes cross-linking of mRNA with proteins S3, S4, S5, S7, S9, S18 and IF-3. Hence, these proteins interact directly with mRNA within the complex 30-S-subunit . mRNA.Addition of IF-3 results in an increase of the rate of complex formation and decrease of its dissociation rate. The addition of IF-3 changes the relative amounts of cross-linked proteins (mainly S3, S4 and S18).Decreasing the temperature from 37 'C to 0 "C not only decelerates the complex formation rate but also changes the relative amount of cross-linked proteins, indicating the influence of the conditions on the structure of the complex.It is well known that prokaryotic messenger RNAs are polycistronic and initiation can take place at the internal sites of the RNA [I]. The selection of initiation sites in messenger RNA depends on RNA conformation ; therefore unfolding of RNA allows the ribosome to interact with additional AUG codons [2,3]. On the other hand, the specificity of initiation is determined by the ribosome itself. For instance, ribosomes of Bacillus stearothermophilus translate predominantly the A protein on RNAs of phages R17 and f 2 [2,4] , whereasEscherichia coli ribosomes translate the coat protein on the same messengers [2].Up to now, the only known molecular interactions involved in initiation complex formation are Shine-Dalgarno interactions, i.e. the complementary binding of the nucleotide sequence near the 3' end of 16-S rRNA and the region in messenger RNA 5'-distal to the initiation codon [S]. However, it has been shown that E. coli ribosomes are able to translate messenger RNAs which do not contain the ShineDalgarno sequence [6]. At the same time eukaryotic ribosomes which possess no polypyrimidine sequence near the 3' end of 1 6 3 RNA [7] also translate RNA from phages giving rise to the same proteins as E. coli ribosomes [8-101. That implies that the initiation complex stabilisation leading to specific interaction between ribosones and messenger RNA depends on a multiplicity of interactions between the respective sites of mRNA and ribosomal components, the relative role of which can change depending on the ribosome and cistron structure. Therefore the structural study of messenger RNA and ribosomal components involved in initiation complex formation is a prerequisite for understanding the principles of initiation. ~~Abhreviarions. Poly(U), poly(uridy1ic acid); poly(s4U), poly(4-thiouridylic acid); IF-3, initiation factor 3 ; rRNA, ribosomal R N A ; 3 0 s . MS2 RNA, complex between 3 0 3 subunits and MS2 RNA; 3 0 3 IF3 . MS2 RNA, complex containing 30-S subunits, IF-3 and MS2 RNA.En;-vmrs. RNase A (EC 3.1.27.5); RNase TI (EC 3.1.27.3); alkaline phosphatase (EC 3.1.3.1); polynucleotide kinase (EC 2.7.1.78). 30-S subunits of E. coli ribosomes form complexes withRNAs from phages in the absence of initiator tRNA [I 1 ...

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[3] Use of in Vitro32P labeling in the sequence analysis of nonradioactive tRNAs

Cited by 442 publications

References 55 publications

The nucleotide sequence of a major glutamine tRNA from rat liver

The nucleotide sequence of a major glutamine tRNA from rat liver

The nucleotide sequence of sheep liver histidine‐tRNA (anticodon Q‐U‐G)

Proteins of the 30‐S Subunit of Escherichia coli Ribosomes which Interact Directly with Natural mRNA

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