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A ribozyme selected from variants of U6 snRNA promotes 2′,5′-branch formation

Published online by Cambridge University Press:  07 February 2001

THOMAS TUSCHL ,
PHILLIP A. SHARP  and
DAVID P. BARTEL
THOMAS TUSCHL
Affiliation:
Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142, USA Present address: Department of Cellular Biochemistry, Max-Planck-Institute for Biophysical Chemistry, D-37070 Göttingen, Germany.
PHILLIP A. SHARP
Affiliation:
Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA
DAVID P. BARTEL
Affiliation:
Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142, USA
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Abstract

In vitro selection was used to sample snRNA-related sequences for ribozyme activities, and several 2′,5′-branch-forming ribozymes were isolated. One such ribozyme is highly dependent upon an 11-nt motif that contains a conserved U6 snRNA sequence (ACAGAGA-box) known to be important for pre-mRNA splicing. The ribozyme reaction is similar to the first step of splicing in that an internal 2′-hydroxyl of an unpaired adenosine attacks at the 5′-phosphate of a guanosine. It differs in that the leaving group is diphosphate rather than a 5′ exon. The finding that lariat formation can be accomplished by a small RNA with sequences related to U6 snRNA indicates that the RNA available in the spliceosome may be involved in RNA-catalyzed branch formation.

Keywords

in vitro selectionRNA catalysissplicing
Type
Research Article
Information
RNA , Volume 7 , Issue 1 , January 2001 , pp. 29 - 43
Copyright
© 2001 RNA Society

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