Nucleic Acids Research, Vol 25, Issue 24 4977-4984, Copyright © 1997 by Oxford University Press
H Ben-Shlomo, A Levitan, O Beja and S Michaeli
We have previously reported the co-purification of a tRNA-like molecule
with the Trypanosoma brucei SRP complex [Beja et al . (1993) Mol. Biochem.
Parasitol . 57, 223-230]. To examine whether the trypanosome SRP has a
unique composition compared with that of other eukaryotes, we analyzed the
7SL RNA and the SRP complex of the monogenetic trypanosomatid Leptomonas
collosoma. The 7SL RNA from L. collosoma was cloned, and its gene was
sequenced. In contrast to T. brucei , two 7SL RNA transcripts were detected
in L.collosoma that originate from a single-copy gene. Using stable cell
lines expressing tagged 7SL RNA, we demonstrate that the tRNAArggene
located 98 bp upstream to the 7SL RNA serves as part of the 7SL RNA
extragenic promoter. The steady-state level of 7SL RNA was found to be
tightly regulated, since the presence of the gene on the multi-copy plasmid
repressed the synthesis of the chromosomal gene. Cell lines carrying
truncated 7SL RNA genes were established and their expression indicated
that domain I is essential for expressing the 7SL RNA. No constructs
carrying portions of the 7SL RNA were expressed, except for a construct
which lacked 23 nt from the 3'end of the RNA. This suggests that 90% of the
7SL RNA molecule is important for its maintenance as a stable small RNA. We
propose that the repression phenomenon may originate from a regulatory
mechanism that coordinates the level of the 7SL RNA by its binding
proteins.
ARTICLES
The trypanosomatid Leptomonas collosoma 7SL RNA gene. Analysis of elements controlling its expression
Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot 76100, Israel.
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