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Mol Cell Biol, June 1998, p. 3431-3444, Vol. 18, No. 6
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.
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
*
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.
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