title: Structural stability of ribosomes subjected to RNase treatment evidenced by dielectric spectroscopy and differential scanning microcalorimetry.
abstract: Previous studies from our laboratory demonstrated the existence of at least two levels of structural complexity in E. coli 70S ribosomes. Ribosomal RNA seems to be principally involved in the overall stability of these structures. In this paper we present an investigation of ribosomes subjected to treatment with RNase. The study is based on both differential scanning microcalorimetry and dielectric spectroscopy. In the thermograms obtained on treated ribosomes only the low temperature peak of the two typical denaturation events observed in native ribosomes, is promptly eliminated by the enzyme treatment. Dielectric spectroscopy measurements carried out on the same samples indicate an alteration of the dielectric behavior previously shown to consist of two subsequent relaxation processes. In fact, only the low frequency relaxation is affected by the treatment. The second one, observed at higher frequency, remains unaltered. The same effect on the dielectric parameters is observed if the ribosome particles are heated and then cooled prior to measurement. These results are consistent with the idea that two different structures are present within the ribosome. One is very stable and withstands both temperature and RNase treatment while the second is promptly abolished by both treatments. Data presented here strongly suggest that the RNA domains exposed to the solvent play a fundamental role in the stability of the 3-D structure of the ribosome particle.
authors.affiliation: INFM-Dipartimento di Fisica, Università La Sapienza, Roma, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Blasi
authors.forename: A
authors.initials: A
authors.lastname: Bonincontro
authors.forename: S
authors.initials: S
authors.lastname: Cinelli
authors.forename: G
authors.initials: G
authors.lastname: Onori
authors.forename: G
authors.initials: G
authors.lastname: Risuleo
mesh_terms: D002152:Calorimetry, Differential Scanning; D011489:Protein Denaturation; D012260:Ribonucleases / Q000494:pharmacology*; D012270:Ribosomes / Q000187:drug effects*; D013057:Spectrum Analysis / Q000379:methods*
chemical_list: D012260:Ribonucleases