Nucleic Acids Research, 2001, Vol. 29, No. 12 2644-2653
© 2001 Oxford University Press
Folding of the group I intron ribozyme from the 26S rRNA gene of Candida albicans
Department of Biotechnology, College of Life Science, Wuhan University, Wuhan 430072, P.R. China, 1Department of Molecular Genetics and Microbiology, University of Medicine and Dentistry, Robert Wood Johnson Medical School, Piscataway, NJ 08854, USA and 2The Cancer Institute of New Jersey, New Brunswick, NJ 08901, USA
Preincubation of the group I intron Ca.LSU from Candida albicans at 37°C in the absence of divalent cations results in partial folding of this intron. This is indicated by increased resistance to T1 ribonuclease cleavage of many G residues in most local helices, including P4-P6, as well as the non-local helix P7, where the G binding site is located. These changes correlate with increased gel mobility and activation of catalysis by precursor RNA containing this intron after preincubation. The presence of divalent cations or spermidine during preincubation results in formation of the predicted helices, as indicated by protection of additional G residues. However, addition of these cations during preincubation of the precursor RNA alters its gel mobility and eliminates the preincubation activation of precursor RNA seen in the absence of cations. These results suggest that, in the presence of divalent cations or spermidine, Ca.LSU folds into a more ordered, stable but misfolded conformation that is less able to convert into the catalytically active form than the ribozyme preincubated without cations. These results indicate that, like the group I intron of Tetrahymena, multiple folding pathways exist for Ca.LSU. However, it appears that the role cations play in the multiple folding pathways leading to the catalytically active form may differ between folding of these two group I introns.
* To whom correspondence should be addressed at: Department of Molecular Genetics and Microbiology, University of Medicine and Dentistry, Robert Wood Johnson Medical School, Piscataway, NJ 08854, USA. Tel: +1 732 235 4795; Fax: +1 732 235 5223; Email: leibowit{at}umdnj.edu
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