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J Biol Chem, Vol. 275, Issue 19, 14767-14776, May 12, 2000
From the Departments of We report the identification of 17 box C/D
fibrillarin-associated small nucleolar RNAs (snoRNAs) from the ancient
eukaryote, Trypanosoma brucei. To systematically isolate
and characterize these snoRNAs, the T. brucei cDNA for
the box C/D snoRNA common protein, fibrillarin, was cloned and
polyclonal antibodies to the recombinant fibrillarin protein were
generated in rabbits. Immunoprecipitations from T. brucei
extracts with the anti-fibrillarin antibodies indicated that this
trypanosomatid has at least 30 fibrillarin-associated snoRNAs. We have
sequenced seventeen of them and designated them TBR for
T.
brucei RNA 1-17. All
of them bear conserved box C, D, C', and D' elements, a hallmark of
fibrillarin-associated snoRNAs in eukaryotes. Fourteen of them are
novel T. brucei snoRNAs. Fifteen bear potential guide regions to mature rRNAs suggesting that they are involved in
2'-O-ribose methylation. Indeed, eight ribose methylations
have been mapped in the rRNA at sites predicted by the snoRNA
sequences. Comparative genomics indicates that six of the seventeen are
the first trypanosome homologs of known yeast and vertebrate
methylation guide snoRNAs. Our results indicate that T. brucei has many fibrillarin-associated box C/D snoRNAs with roles
in 2'-O-ribose methylation of rRNA and that the mechanism
for targeting the nucleotide to be methylated at the fifth nucleotide
upstream of box D or D' originated in early eukaryotes.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF168719-AF168736.
Fibrillarin-associated Box C/D Small Nucleolar RNAs in
Trypanosoma brucei
SEQUENCE CONSERVATION AND IMPLICATIONS FOR
2'-O-RIBOSE METHYLATION OF rRNA*
,
**
Therapeutic Radiology,
§ Cell Biology, and Genetics, Yale School of
Medicine, New Haven, Connecticut 06520-8040 and the
¶ Department of Genetics, Washington University School of
Medicine, St. Louis, Missouri 63110
*
This work was supported by a grant from the Council for
Tobacco Research (to S. J. B.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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