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Rapid global structure determination of large RNA and RNA complexes using NMR and
small-angle X-ray scattering
#MMPMID20554045
Wang YX
; Zuo X
; Wang J
; Yu P
; Butcher SE
Methods
2010[Oct]; 52
(2
): 180-91
PMID20554045
show ga
Among the greatest advances in biology today are the discoveries of various roles
played by RNA in biological processes. However, despite significant advances in
RNA structure determination using X-ray crystallography [1] and solution NMR
[2-4], the number of bona fide RNA structures is very limited, in comparison with
the growing number of known functional RNAs. This is because of great difficulty
in growing crystals or/and obtaining phase information, and severe size
constraints on structure determination by solution NMR spectroscopy. Clearly,
there is an acute need for new methodologies for RNA structure determination. The
prevailing approach for structure determination of RNA in solution is a
"bottom-up" approach that was basically transplanted from the approach used for
determining protein structures, despite vast differences in both structural
features and chemical compositions between these two types of biomacromolecules.
In this chapter, we describe a new method, which has been reported recently, for
rapid global structure determination of RNAs using solution-based NMR
spectroscopy and small-angle X-ray scattering. The method treats duplexes as
major building blocks of RNA structures. By determining the global orientations
of the duplexes and the overall shape, the global structure of an RNA can be
constructed and further regularized using Xplor-NIH. The utility of the method
was demonstrated in global structure determination of two RNAs, a 71-nt and
102-nt RNAs with an estimated backbone RMSD ?3.0Å. The global structure opens
door to high-resolution structure determination in solution.
|*Scattering, Small Angle
[MESH]
|Models, Molecular
[MESH]
|Nuclear Magnetic Resonance, Biomolecular/*methods
[MESH]
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