Mapping of yeast tRNAs by two‐dimensional electrophoresis on polyacrylamide gels

Fradin, Anny; Gruhl, Hartmut; Feldmann, Horst

doi:10.1016/0014-5793(75)80485-6

Cited by 136 publications

(46 citation statements)

References 16 publications

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“…5SS and 5Ssom RNAs were resolved using an acrylamide gel system adapted from that of Fradin et al (1975). Both stacking and resolving gels contain 4 M urea and have acrylamide:bis ratios of 27:1.…”

Section: Methodsmentioning

confidence: 99%

Cytoplasmic regulation of 5S RNA genes in nuclear-transplant embryos.

Wakefield

Gurdon

1983

The EMBO Journal

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In the normal development of Xenopus laevis, genes for oocyte-type and somatic-type 5S RNAs are both expressed in late blastulae. Estimates of rates of synthesis indicate that the oocyte-type genes (5S°O9 undergo at least a 100-fold reduction in transcriptional activity between the end of oogenesis and the late blastula stage, and at least a further 20-fold reduction during gastrulation. When neurula nuclei, with inactive 5SO°genes, were transplanted to enucleated eggs, the resulting nuclear-transplant embryos showed a reactivation of 550c genes in blastulae and a subsequent inactivation after this stage. This effect is not explicable by a differential stability of the two types of 5S RNA. We conclude that egg or early embryo cytoplasm must contain components which can continuously regulate 5S gene expression, and that the mechanism by which 5S°0c genes are developmentally inactivated does not persist through mitosis in early embryos. These results have been obtained by a new procedure in which oocyte-and somatic-type 5S RNAs are separated in a 4 M urea-1507o acrylamide gel.

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Section: Methodsmentioning

confidence: 99%

Cytoplasmic regulation of 5S RNA genes in nuclear-transplant embryos.

Wakefield

Gurdon

1983

The EMBO Journal

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“…Protein bands were detected by either Coomassie Blue or silver staining according to the manufacturer's recommendations. tRNAs were analysed on 10% polyacrylamide gels containing 7 M urea [25] and detected by staining with Stains AllR (Serva; [26]). …”

Section: Gel Electrophoretic Analysis Of Proteins and Rnamentioning

confidence: 99%

Discrimination against misacylated tRNA by chloroplast elongation factor Tu

Stanzel

Schön

Sprinzl

1994

European Journal of Biochemistry

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Chloroplast elongation factor Tu was purified from Pisum sativum and the binding properties of glutamylated chloroplast tRNAs were studied by gel‐permeation chromatography. Whereas chloroplast Glu‐tRNAGlu is efficiently bound by this factor, the misacylated Glu‐tRNAGin does not interact with chloroplast elongation factor Tu · GTP and is thus efficiently excluded from protein synthesis. Comparison with the behaviour of Escherichia coli elongation factor Tu · GTP shows that this factor, which is not confronted with the in vivo misacylation phenomenon of organelles, binds both Glu‐tRNAGlu and Glu‐tRNAGln from chloroplasts with approximately equal efficiency.

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“…Recently, we devised a method for mapping tRNAs by the use of two-dimensional electrophoresis on polyacrylamide gels [22]. This technique was applied to locust mitochondrial tRNA in order to get an estimate about the number of individual tRNA species and to prove the existence of isoacceptors.…”

Section: Mapping Of Mitochondrial Trna Species On Polyacrylamide Gelsmentioning

confidence: 99%

“…Electrophoresis of locust tRNAs on polyacrylamide gels. (a) 10% gel, technical details were as described [22]. A: mitochondrial tRNA.…”

Section: Mapping Of Mitochondrial Trna Species On Polyacrylamide Gelsmentioning

confidence: 99%

“…The homogenate was centrifuged for 15 min at 12 000 × g and then recentrifuged for 2 h at 226 000 × g. The upper half of the supernatant was treated as described above. The conditions for aminoacylation of the tRNAs mere those described [22]. 3H-or 14C-labeled amino acids with high specific activities (The Radiochemical Centre, Amersham) were used.…”

Section: Mapping Of Mitochondrial Trna Species On Polyacrylamide Gelsmentioning

confidence: 99%

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