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Volume 272, Number 35, Issue of August 29, 1997 pp. 21989-21993
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

Formation of 2,3-Cyclic Phosphates at the 3 End of Human U6 Small Nuclear RNA in Vitro
IDENTIFICATION OF 2,3-CYCLIC PHOSPHATES AT THE 3 ENDS OF HUMAN SIGNAL RECOGNITION PARTICLE AND MITOCHONDRIAL RNA PROCESSING RNAs

(Received for publication, April 3, 1997, and in revised form, June 25, 1997)

From Baylor College of Medicine, Department of Pharmacology, Houston, Texas 77030

Approximately 90% of human U6 small nuclear RNA (snRNA) contains uridine cyclic phosphate (U>p) at its 3-end (Lund, E., and Dahlberg, J. E. (1992) Science 255, 327-330). We studied the formation of U>p at the 3 end of human U6 snRNA using an in vitro system where uridylic acid residues are added from UTP precursor and U>p is formed. Analysis of U6 snRNAs with varying number of uridylic acid residues showed that each of these species contains U>p where the phosphate originated from -phosphate of UTP precursor. The cyclic phosphate formation occurred on U6 snRNA in extracts where essential spliceosomal snRNAs were specifically degraded, thereby indicating that U>p formation is not coupled to pre-mRNA splicing. A subpopulation of human signal recognition particle and mitochondrial RNA processing RNAs isolated from HeLa cells also contained cyclic phosphates at their 3 ends. These data suggest that U>p in U6 snRNA is unlikely to be related to its participation in splicing of pre-mRNAs. It appears that cyclic phosphate is an intermediate product in the metabolism of these small RNAs.

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