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Mol Cell Biol, June 1998, p. 3431-3444, Vol. 18, No. 6
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Functional Mapping of the U3 Small Nucleolar RNA from the Yeast Saccharomyces cerevisiae

Dmitry A. Samarsky and Maurille J. Fournier*

Department of Biochemistry and Molecular Biology, Program in Molecular and Cellular Biology, University of Massachusetts, Amherst, Massachusetts 01003

Received 22 December 1997/Returned for modification 22 January 1998/Accepted 5 March 1998

The U3 small nucleolar RNA participates in early events of eukaryotic pre-rRNA cleavage and is essential for formation of 18S rRNA. Using an in vivo system, we have developed a functional map of the U3 small nucleolar RNA from Saccharomyces cerevisiae. The test strain features a galactose-dependent U3 gene in the chromosome and a plasmid-encoded allele with a unique hybridization tag. Effects of mutations on U3 production were analyzed by evaluating RNA levels in cells grown on galactose medium, and effects on U3 function were assessed by growing cells on glucose medium. The major findings are as follows: (i) boxes C' and D and flanking helices are critical for U3 accumulation; (ii) boxes B and C are not essential for U3 production but are important for function, most likely due to binding of a trans-acting factor(s); (iii) the 5' portion of U3 is required for function but not stability; and, (iv) strikingly, the nonconserved hairpins 2, 3, and 4, which account for 50% of the molecule, are not required for accumulation or function.

* Corresponding author. Mailing address: Department of Biochemistry and Molecular Biology, Lederle GRC Tower B, University of Massachusetts, Box 34505, Amherst, MA 01003-4505. Phone: (413) 545-2732. Fax: (413) 545-3291. E-mail: 4nier{at}biochem.umass.edu.

Mol Cell Biol, June 1998, p. 3431-3444, Vol. 18, No. 6
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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Copyright © 1998 by the American Society for Microbiology. All rights reserved.