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Bulge-induced bends in RNA: quantification by transient electric birefringence.
J Mol Biol. 1995 Mar 31; 247(3):486-500.JM

Abstract

Bulges represent one of the most common non-helical elements in RNA, often displaying a strong degree of phylogenetic conservation, both in location and sequence, within larger RNA molecules. Thus, knowledge of the conformation and flexibility of RNA bulges is an important prerequisite for understanding the rules governing the formation of tertiary structure within the larger molecules. In the current investigation, the magnitudes of the bends induced in a 148 base-pair duplex RNA molecule by single, centrally located bulges of varying size (n = 1 to 6) and base composition (An and Un series) have been determined through the use of transient electric birefringence (TEB). The TEB approach is highly sensitive to the changes in the global shape of RNA (or DNA) helices that accompany the introduction of points of bending or flexibility near the center of the helix. In the current instance, bulge angles deduced from TEB measurements ranged from approximately 7 degrees to approximately 93 degrees, with the angle increasing with increasing n for both An and Un series. For both An and Un series in the absence of Mg2+, the angle increment per added nucleotide varied from approximately 20 degrees to approximately 8 degrees as n increased from 1 to 6. These angle increments remained unchanged for the An series in the presence of Mg2+; however, the angle increments for the Un series were reduced by a factor of 2 for all values of n. Thus the current observations have identified structural transitions in one of the simplest non-helical elements in RNA, transitions that are dependent on both sequence and counterion valence. Finally, the measured bend angles are strongly correlated with the degree of reduction in electrophoretic mobility of bulge-containing RNA helices. The observed correlation was used to obtain a semi-empirical relationship between bend angle and mobility in order that additional angles might be assigned, by interpolation, through the use of gel data alone.

Authors+Show Affiliations

Department of Biochemistry, Biophysics and Genetics, University of Colorado Health Sciences Center, Denver, USA.No affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

7536250

Citation

Zacharias, M, and P J. Hagerman. "Bulge-induced Bends in RNA: Quantification By Transient Electric Birefringence." Journal of Molecular Biology, vol. 247, no. 3, 1995, pp. 486-500.
Zacharias M, Hagerman PJ. Bulge-induced bends in RNA: quantification by transient electric birefringence. J Mol Biol. 1995;247(3):486-500.
Zacharias, M., & Hagerman, P. J. (1995). Bulge-induced bends in RNA: quantification by transient electric birefringence. Journal of Molecular Biology, 247(3), 486-500.
Zacharias M, Hagerman PJ. Bulge-induced Bends in RNA: Quantification By Transient Electric Birefringence. J Mol Biol. 1995 Mar 31;247(3):486-500. PubMed PMID: 7536250.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Bulge-induced bends in RNA: quantification by transient electric birefringence. AU - Zacharias,M, AU - Hagerman,P J, PY - 1995/3/31/pubmed PY - 1995/3/31/medline PY - 1995/3/31/entrez SP - 486 EP - 500 JF - Journal of molecular biology JO - J Mol Biol VL - 247 IS - 3 N2 - Bulges represent one of the most common non-helical elements in RNA, often displaying a strong degree of phylogenetic conservation, both in location and sequence, within larger RNA molecules. Thus, knowledge of the conformation and flexibility of RNA bulges is an important prerequisite for understanding the rules governing the formation of tertiary structure within the larger molecules. In the current investigation, the magnitudes of the bends induced in a 148 base-pair duplex RNA molecule by single, centrally located bulges of varying size (n = 1 to 6) and base composition (An and Un series) have been determined through the use of transient electric birefringence (TEB). The TEB approach is highly sensitive to the changes in the global shape of RNA (or DNA) helices that accompany the introduction of points of bending or flexibility near the center of the helix. In the current instance, bulge angles deduced from TEB measurements ranged from approximately 7 degrees to approximately 93 degrees, with the angle increasing with increasing n for both An and Un series. For both An and Un series in the absence of Mg2+, the angle increment per added nucleotide varied from approximately 20 degrees to approximately 8 degrees as n increased from 1 to 6. These angle increments remained unchanged for the An series in the presence of Mg2+; however, the angle increments for the Un series were reduced by a factor of 2 for all values of n. Thus the current observations have identified structural transitions in one of the simplest non-helical elements in RNA, transitions that are dependent on both sequence and counterion valence. Finally, the measured bend angles are strongly correlated with the degree of reduction in electrophoretic mobility of bulge-containing RNA helices. The observed correlation was used to obtain a semi-empirical relationship between bend angle and mobility in order that additional angles might be assigned, by interpolation, through the use of gel data alone. SN - 0022-2836 UR - https://www.unboundmedicine.com/medline/citation/7536250/Bulge_induced_bends_in_RNA:_quantification_by_transient_electric_birefringence_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-2836(85)70155-6 DB - PRIME DP - Unbound Medicine ER -