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J. Biol. Chem., Vol. 266, Issue 34, 22792-22795, Dec, 1991
GJ Hannon, PA Maroney and TW Nilsen
In nematodes, a fraction of mRNAs acquires a common 22-nucleotide 5'-
terminal spliced leader sequence via a trans-splicing reaction. The same
premessenger RNAs which receive the spliced leader are also processed by
conventional cis-splicing. Whole cell extracts prepared from synchronous
embryos of the parasitic nematode Ascaris lumbricoides catalyze both cis-
and trans-splicing. We have used this cell-free system and
oligodeoxynucleotide directed RNase H digestion to assess the U small
nuclear RNA requirements for nematode cis- and trans- splicing. These
experiments indicated that both cis- and trans-splicing require intact U2
and U4/U6 small nuclear ribonucleoproteins (snRNPs). However, whereas
cis-splicing displays the expected requirement for an intact U1 snRNP,
trans-splicing is unaffected when approximately 90% of U1 snRNP is
degraded. These results suggest that 5' splice site identification differs
in nematode cis- and trans-splicing.
U small nuclear ribonucleoprotein requirements for nematode cis- and trans-splicing in vitro
Department of Molecular Biology and Microbiology, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106-4960.
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