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J Biol Chem, Vol. 274, Issue 27, 19361-19367, July 2, 1999
From the Department of Microbiology and Immunology, UCLA School of
Medicine, Los Angeles, California 90095-1747
A 39-nucleotide leader is
trans-spliced onto all trypanosome nuclear mRNAs. The
precursor spliced leader RNA was tested for trans-splicing
function in vivo by mutating the intron. We report that in
Leishmania tarentolae spliced leader RNA 5' modification is
influenced by the primary sequence of stem-loop II, the Sm-binding site, and the secondary structure of stem-loop III. The sequence of
stem-loop II was found to be important for cap 4 formation and
splicing. As in Ascaris, mutagenesis of the bulge
nucleotide in stem-loop II was detrimental to
trans-splicing. Because restoration of the L. tarentolae stem-loop II structure was not sufficient to restore
splicing, this result contrasts the findings in the kinetoplastid
Leptomonas, where mutations that restored stem-loop II
structure supported splicing. Methylation of the cap 4 structure and
splicing was also dependent on both the Sm-binding site and the
structure of stem-loop III and was inhibited by incomplete 3' end
processing. The critical nature of the L. tarentolae
Sm-binding site is consistent with its essential role in the
Ascaris spliced leader RNA, whereas in
Leptomonas mutation of the Sm-binding site and deletion of
stem-loop III did not affect trans-splicing. A pathway for
Leishmania spliced leader RNA processing and maturation is proposed.
The Role of Intron Structures in trans-Splicing and
Cap 4 Formation for the Leishmania Spliced Leader RNA
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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