title: Evaluation of parameters critical for observing nucleic acids inside living Xenopus laevis oocytes by in-cell NMR spectroscopy.
abstract: In-cell NMR spectroscopy of proteins in different cellular environments is a well-established technique that, however, has not been applied to nucleic acids so far. Here, we show that isotopically labeled DNA and RNA can be observed inside the eukaryotic environment of Xenopus laevis oocytes by in-cell NMR spectroscopy. One limiting factor for the observation of nucleic acids in Xenopus oocytes is their reduced stability. We demonstrate that chemical modification of DNA and RNA can protect them from degradation and can significantly enhance their lifetime. Finally, we show that the imino region of the NMR spectrum is devoid of any oocyte background signals enabling the detection even of isotopically nonlabeled molecules.
authors.affiliation: Institute of Biophysical Chemistry, Goethe-University, Max-von-Laue Str. 9, 60438 Frankfurt am Main, Germany.
authors.forename: Robert
authors.initials: R
authors.lastname: Hänsel
authors.forename: Silvie
authors.initials: S
authors.lastname: Foldynová-Trantírková
authors.forename: Frank
authors.initials: F
authors.lastname: Löhr
authors.forename: Janina
authors.initials: J
authors.lastname: Buck
authors.forename: Eva
authors.initials: E
authors.lastname: Bongartz
authors.forename: Ernst
authors.initials: E
authors.lastname: Bamberg
authors.forename: Harald
authors.initials: H
authors.lastname: Schwalbe
authors.forename: Volker
authors.initials: V
authors.lastname: Dötsch
authors.forename: Lukás
authors.initials: L
authors.lastname: Trantírek
mesh_terms: D000818:Animals; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D009696:Nucleic Acids / Q000378:metabolism*; D009841:Oligonucleotides; D009865:Oocytes / Q000378:metabolism*; D012091:Repetitive Sequences, Nucleic Acid; D014982:Xenopus laevis
chemical_list: D009696:Nucleic Acids; D009841:Oligonucleotides