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A chicken embryo protein related to the mammalian DEAD box protein p68 is tightly associated with the highly purified protein-RNA complex of 5-MeC-DNA glycosylase

Jost, Jean-Pierre ; Schwarz, Steffen ; Hess, Daniel ; Angliker, Herbert ; Fuller-Pace, Frances V. ; Stahl, Hans ; Thiry, Stéphane ; Siegmann, Michel

In: Nucleic Acids Research, 1999, vol. 27, no. 16, p. 3245-3252

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    Titre
    • A chicken embryo protein related to the mammalian DEAD box protein p68 is tightly associated with the highly purified protein-RNA complex of 5-MeC-DNA glycosylase
    Auteur
    • Jost, Jean-Pierre. Friedrich Miescher-Institute, PO Box 2543, CH-4002 Basel, Switzerland, Dundee DD1 9SY, UK, D-66421 Homburg/Saar, Germany
    • Schwarz, Steffen. Friedrich Miescher-Institute, PO Box 2543, CH-4002 Basel, Switzerland, Dundee DD1 9SY, UK, D-66421 Homburg/Saar, Germany
    • Hess, Daniel. Friedrich Miescher-Institute, PO Box 2543, CH-4002 Basel, Switzerland, Dundee DD1 9SY, UK, D-66421 Homburg/Saar, Germany
    • Angliker, Herbert. Friedrich Miescher-Institute, PO Box 2543, CH-4002 Basel, Switzerland, Dundee DD1 9SY, UK, D-66421 Homburg/Saar, Germany
    • Fuller-Pace, Frances V.. Department of Molecular and Cellular Pathology, University of Dundee, Ninewells Medical School, Dundee DD1 9SY, UK
    • Stahl, Hans. Medizinische Biochemie und Molekularbiologie, Universität des Saarlandes, D-66421 Homburg/Saar, Germany
    • Thiry, Stéphane. Friedrich Miescher-Institute, PO Box 2543, CH-4002 Basel, Switzerland, Dundee DD1 9SY, UK, D-66421 Homburg/Saar, Germany
    • Siegmann, Michel. Friedrich Miescher-Institute, PO Box 2543, CH-4002 Basel, Switzerland, Dundee DD1 9SY, UK, D-66421 Homburg/Saar, Germany
    Type de document
    • Postprint
    Langue
    • Anglais
    Publié dans
    • Nucleic Acids Research, 1999, vol. 27, no. 16, p. 3245-3252. Oxford University Press
    Autre version électronique
    Classification
    • Santé
    Identifiant OAI-PMH
    • oai:doc.rero.ch:291554
    Summary
    We have shown previously that DNA demethylation by chick embryo 5-methylcytosine (5-MeC)-DNA glycosylase needs both protein and RNA. Amino acid sequences of nine peptides derived from a highly purified 5-MeC-DNA glycosylase complex were identified by Nanoelectrospray ionisation mass spec-trometry to be identical to the mammalian nuclear DEAD box protein p68 RNA helicase. Antibodies directed against human p68 helicase cross-reacted with the purified 5-MeC-DNA glycosylase complex and immunoprecipitated the glycosylase activity. A 2690 bp cDNA coding for the chicken homologue of mammalian p68 was isolated and sequenced. Its derived amino acid sequence is almost identical to the human p68 DEAD box protein up to amino acid position 473 (from a total of 595). This sequence contains all the essential conserved motifs from the DEAD box proteins which are the ATPase, RNA unwinding and RNA binding motifs. The rest of the 122 amino acids in the C-terminal region rather diverge from the human p68 RNA helicase sequence. The recombinant chicken DEAD box protein expressed in Escherichia coli cross-reacts with the same p68 antibodies as the purified chicken embryo 5-MeC-DNA glycosylase complex. The recombinant protein has an RNA-dependent ATPase and an ATP-dependent helicase activity. However, in the presence or absence of RNA the recombinant protein had no 5-MeC-DNA glycosylase activity. In situ hybridisation of 5 day-old chicken embryos with antisense probes of the chicken DEAD box protein shows a high abundance of its transcripts in differentiating embryonic tissues