Publications
1992
Heitzler J., Marechal-Drouard L., Dirheimer G., Keith G.
Use of a dot blot hybridization method for identification of pure tRNA species on different membranes Journal Article
In: Biochim Biophys Acta-Gene Regul Mech, vol. 1129, no. 3, pp. 273-7, 1992, (0006-3002 Journal Article).
Abstract | BibTeX | Tags: *Membranes, Acid, Artificial, Autoradiography, cerevisiae/genetics, Fungal/genetics, Gov't, Hybridization, Met/genetics, Non-U.S., Nucleic, RNA, Saccharomyces, Support, Transfer, Transfer/*genetics
@article{,
title = {Use of a dot blot hybridization method for identification of pure tRNA species on different membranes},
author = { J. Heitzler and L. Marechal-Drouard and G. Dirheimer and G. Keith},
year = {1992},
date = {1992-01-01},
journal = {Biochim Biophys Acta-Gene Regul Mech},
volume = {1129},
number = {3},
pages = {273-7},
abstract = {The characterization of a tRNA in purification procedures usually involves aminoacylation assays but recently, the hybridization by dot blot with specific oligonucleotides as probes has been used for the tRNA identification. We present here an optimization of a dot blot hybridization method for the tRNA detection by comparing the efficiency of eight different nylon membranes. Neutral 0.22 microns porosity membranes (Nytran, Biodine A) give the best detection efficiency when small quantities of material (less than 40 ng of tRNA) are dotted on filter; by contrast, neutral 0.45 microns porosity membranes (such as Hybond N) are the most efficient when larger quantities of tRNA are dotted on the filter. The described technique allows to detect less than 20 pg of a pure tRNA species. Its use in the identification of Saccharomyces cerevisiae initiator tRNA(Met) in counter-current distribution fractions is shown.},
note = {0006-3002
Journal Article},
keywords = {*Membranes, Acid, Artificial, Autoradiography, cerevisiae/genetics, Fungal/genetics, Gov't, Hybridization, Met/genetics, Non-U.S., Nucleic, RNA, Saccharomyces, Support, Transfer, Transfer/*genetics},
pubstate = {published},
tppubtype = {article}
}
Nothwang H. G., Coux O., Keith G., Silva-Pereira I., Scherrer K.
The major RNA in prosomes of HeLa cells and duck erythroblasts is tRNA(Lys,3) Journal Article
In: Nucleic Acids Res, vol. 20, no. 8, pp. 1959-65, 1992, (0305-1048 Journal Article).
Abstract | BibTeX | Tags: Animals, Base, Blotting, Cells, Data, Ducks, effects, Electrophoresis, Erythroblasts, Gel, Gov't, Hela, Human, Lys/*analysis/metabolism, Molecular, Non-U.S., Northern, Nucleotidyltransferases/metabolism, Ribonucleoproteins/*chemistry/drug, RNA, Sequence, Support, Transfer, Two-Dimensional, Zinc/pharmacology
@article{,
title = {The major RNA in prosomes of HeLa cells and duck erythroblasts is tRNA(Lys,3)},
author = { H. G. Nothwang and O. Coux and G. Keith and I. Silva-Pereira and K. Scherrer},
year = {1992},
date = {1992-01-01},
journal = {Nucleic Acids Res},
volume = {20},
number = {8},
pages = {1959-65},
abstract = {Two-dimensional gel electrophoresis of HeLa cell prosomal RNAs, 3'-end labeled by RNA ligase, revealed one prominent spot. Determination of a partial sequence at the 3'-end indicated full homology to the 18 nucleotides at the 3'-end of tRNA(Lys,3) from rabbit, the bovine and the human species. An oligonucleotide complementary to the 3'-end of tRNA(Lys,3) hybridized on Northern blots with prosomal RNA from both HeLa cells and duck erythroblasts. In two-dimensional PAGE, the major pRNA of HeLa cells co-migrated with bovine tRNA(Lys,3). Reconstitution of the CCA 3'-end of RNA from both human and duck prosomes, by tRNA-nucleotidyl-transferase, confirmed the tRNA character of this type of RNA. Furthermore, it revealed at least one additional tRNA band about 85 nt long among the prosomal RNA from both species. Finally, confirming an original property of prosomal RNA, we show that in vitro synthesized tRNA(Lys,3) hybridizes stably to duck globin mRNA, and to poly(A)(+)- and poly(A)(-)-RNA from HeLa cells.},
note = {0305-1048
Journal Article},
keywords = {Animals, Base, Blotting, Cells, Data, Ducks, effects, Electrophoresis, Erythroblasts, Gel, Gov't, Hela, Human, Lys/*analysis/metabolism, Molecular, Non-U.S., Northern, Nucleotidyltransferases/metabolism, Ribonucleoproteins/*chemistry/drug, RNA, Sequence, Support, Transfer, Two-Dimensional, Zinc/pharmacology},
pubstate = {published},
tppubtype = {article}
}
Przykorska A., el Adlouni C., Keith G., Szarkowski J. W., Dirheimer G.
Structural specificity of Rn nuclease I as probed on yeast tRNA(Phe) and tRNA(Asp) Journal Article
In: Nucleic Acids Res, vol. 20, no. 4, pp. 659-63, 1992, (0305-1048 Journal Article).
Abstract | BibTeX | Tags: Acid, Asp/chemistry/genetics/*metabolism, Base, cereale, Composition, Conformation, Data, Gov't, Molecular, Non-U.S., Nucleic, Pancreatic/*metabolism, Phe/chemistry/genetics/*metabolism, Ribonuclease, RNA, Secale, Sequence, Specificity, Substrate, Support, Transfer, Yeasts/genetics
@article{,
title = {Structural specificity of Rn nuclease I as probed on yeast tRNA(Phe) and tRNA(Asp)},
author = { A. Przykorska and C. el Adlouni and G. Keith and J. W. Szarkowski and G. Dirheimer},
year = {1992},
date = {1992-01-01},
journal = {Nucleic Acids Res},
volume = {20},
number = {4},
pages = {659-63},
abstract = {A single-strand-specific nuclease from rye germ (Rn nuclease I) was characterized as a tool for secondary and tertiary structure investigation of RNAs. To test the procedure, yeast tRNA(Phe) and tRNA(Asp) for which the tertiary structures are known, as well as the 3'-half of tRNA(Asp) were used as substrates. In tRNA(Phe) the nuclease introduced main primary cuts at positions U33 and A35 of the anticodon loop and G18 and G19 of the D loop. No primary cuts were observed within the double stranded stems. In tRNA(Asp) the main cuts occurred at positions U33, G34, U35, C36 of the anticodon loop and G18 and C20:1 positions in the D loop. No cuts were observed in the T loop in intact tRNA(Asp) but strong primary cleavages occurred at positions psi 55, C56, A57 within that loop in the absence of the tertiary interactions between T and D loops (use of 3'-half tRNA(Asp)). These results show that Rn nuclease I is specific for exposed single-stranded regions.},
note = {0305-1048
Journal Article},
keywords = {Acid, Asp/chemistry/genetics/*metabolism, Base, cereale, Composition, Conformation, Data, Gov't, Molecular, Non-U.S., Nucleic, Pancreatic/*metabolism, Phe/chemistry/genetics/*metabolism, Ribonuclease, RNA, Secale, Sequence, Specificity, Substrate, Support, Transfer, Yeasts/genetics},
pubstate = {published},
tppubtype = {article}
}