NDLI: Crystallization of ribozymes and small RNA motifs by a sparse matrix approach.

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Crystallization of ribozymes and small RNA motifs by a sparse matrix approach.

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Crystallization of ribozymes and small RNA motifs by a sparse matrix approach.

Content Provider	PubMed Central
Author	Doudna, J. A. Grosshans, C. Gooding, A. Kundrot, C. E.
Abstract	The three-dimensional structures of RNA enzymes form catalytic centers that include specific substrate binding sites. High-resolution determination of these and other RNA structures is essential for a detailed understanding of the function of RNA in biological systems. The crystal structures of only a few RNA molecules are currently known. These include tRNAs, which were produced in vivo and contained modified bases, and short oligonucleotide duplexes lacking tertiary interactions. Here we report that a number of different RNA molecules of 4-50 kDa, all synthesized in vitro, have been crystallized. A highly successful method for the growth of RNA crystals based on previously reported conditions for tRNA crystallization is presented. This method is rapid and economical, typically requiring 1.1 mg of RNA to set up an experiment and 2 weeks to complete the observations. Using this technique, we have obtained crystals of 8 of 10 different RNA molecules tested, ranging in size from a dodecamer duplex to a 208-nucleotide catalytic intron. Several of these crystal forms diffract to high resolution; in one case, we have collected a 2.8-A native data set for a 160-nucleotide domain of the group I self-splicing intron from Tetrahymena thermophila. The solution of these RNA structures should reveal aspects of tertiary structure that relate to RNA function and catalytic mechanisms.
Starting Page	7829
File Format	PDF
ISSN	10916490
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	16
Volume Number	90
Language	English
Publisher Date	1993-08-15
Access Restriction	Open
Subject Keyword	Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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