Organization and transient expression of the gene for human U11 snRNA.
The nucleotide sequence of U11 small nuclear RNA, a minor U RNA from HeLa cells, was determined. Computer analysis of the sequence (135 residues) predicts two strong hairpin loops which are separated by seventeen nucleotides containing an Sm binding site (AAUUUUUUGG). A synthetic gene was constructed in which the coding region of U11 RNA is under the control of a T7 promoter. This vector can be used to produce U11 RNA in vitro. Southern hybridization and PCR analysis of HeLa genomic DNA suggest that U11 RNA is encoded by a single copy gene, and that at least three genomic regions could be U11 RNA pseudogenes. A HeLa genomic copy of a U11 gene was isolated by inverted PCR. This gene contains the U11 RNA coding sequence and several sequence elements unique for the U RNA genes. These include a Distal Sequence Element (DSE, ATTTGCATA) present between positions -215 and -223 relative to the start of transcription; a Proximal Sequence Element (PSE, TTCACCTTTACCAAAAATG) located between positions -43 and -63; and a 3' box (GTTAGGCGAAATATTA) between positions + 150 and + 166. Transfection of HeLa cells with this gene revealed that it is functioning in vivo and can produce U11 RNA.
Suter-Crazzolara
C
C
Department of Cell Biology, University of Basel, Switzerland.
Keller
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W
D001483:Base Sequence; D003001:Cloning, Molecular; D015870:Gene Expression; D015698:Genomic Library; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D011544:Pseudogenes; D012342:RNA, Small Nuclear / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection
D012342:RNA, Small Nuclear