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Papers In Press, published online ahead of print November 7, 2001
Department of Biochemistry, University of Washington, Seattle, WA 98195-7350
Corresponding Author: amweiner{at}u.washington.edu
The CCA-adding enzyme builds and repairs the 3' terminus of tRNA. Approximately 65% of mature human U2 snRNA ends in 3' terminal CCA, as do all mature tRNAs; the other 35% ends in 3' CC or possibly 3' C. The 3' terminal A of U2 snRNA cannot be encoded, because the 3' end of the U2 snRNA coding region is CC/CC where the slash indicates the last encoded nucleotide. The first detectable U2 snRNA precursor contains 10 to 16 extra 3' nucleotides that are removed by one or more 3' exonucleases. Thus if 3' exonuclease activity removes the encoded 3' CC during U2 snRNA maturation, as appears to be the case in vitro, the cell may need to build or rebuild the 3' terminal A, CA, or CCA of U2 snRNA. We asked whether homologous and heterologous class I and class II CCA-adding enzymes could add 3' terminal A, CA, or CCA to human U2 snRNA lacking 3' terminal A, CA, or CCA. The naked U2 snRNAs were good substrates for the human CCA-adding enzyme, but inactive with the E. coli enzyme; activity was also observed on native U2 snRNPs. We suggest that the 3' stem/loop of U2 snRNA resembles a tRNA minihelix, the smallest efficient substrate for class I and II CCA-adding enzymes, and that CCA addition to U2 snRNA may take place in vivo after snRNP assembly has begun.
J. Biol. Chem, 10.1074/jbc.M109559200
Submitted on October 3, 2001
Revised on November 7, 2001
Accepted on November 6, 2001
U2 snRNA is a substrate for the CCA-adding enzyme (tRNA nucleotidyltransferase)
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