NMR reveals the absence of hydrogen bonding in adjacent UU and AG mismatches in an isolated internal loop from ribosomal RNA.
NMR studies provide insights into structural features of internal loops. These insights can be combined with thermodynamic studies to generate models for predicting structure and energetics. The tandem mismatch internal loop, 5'GUGG3'(3'CUAC5'), has been studied by NMR. The NMR structure reveals an internal loop with no hydrogen bonding between the loop bases and with the G in the AG mismatch flipped out of the helix. The sequence of this internal loop is highly conserved in rRNA. The loop is located in the large ribosomal subunit and is part of a conserved 58-nt fragment that is the binding domain of ribosomal protein L11. Structural comparisons between variants of this internal loop in crystal structures of the 58-nt domain complexed with L11 protein and of the large ribosomal subunit (LSU) suggest that this thermodynamically destabilizing internal loop is partially preorganized for tertiary interactions and for binding L11. A model for predicting the base pairing and free energy of 2 x 2 nucleotide internal loops with a purine-purine mismatch next to a pyrimidine-pyrimidine mismatch is proposed on the basis of the present NMR structure and previously reported thermodynamics.
Shankar
Neelaabh
N
Department of Biochemistry and Biophysics, University of Rochester, Rochester, New York 14642, USA.
Xia
Tianbing
T
Kennedy
Scott D
SD
Krugh
Thomas R
TR
Mathews
David H
DH
Turner
Douglas H
DH
D000818:Animals; D020137:Base Pair Mismatch; D001483:Base Sequence; D017124:Conserved Sequence; D016829:Giardia lamblia / Q000235:genetics; D006860:Hydrogen Bonding; D008954:Models, Biological; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D009690:Nucleic Acid Conformation; D012335:RNA, Ribosomal / Q000737:chemistry*; D054658:Ribosome Subunits, Large / Q000235:genetics*; D016945:Sulfolobus acidocaldarius / Q000235:genetics; D014982:Xenopus laevis / Q000235:genetics; D003313:Zea mays / Q000235:genetics
D012335:RNA, Ribosomal