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MOLECULAR AND CELLULAR BIOLOGY, JUlY 1985, p. 1560-1570 Vol. 5, No. 7 0270-7306/85/071560-11$02.00/0
 

Summary: MOLECULAR AND CELLULAR BIOLOGY, JUlY 1985, p. 1560-1570 Vol. 5, No. 7
0270-7306/85/071560-11$02.00/0
Copyright ęD 1985, American Society for Microbiology
Orientation-Dependent Transcriptional Activator Upstream of a
Human U2 snRNA Gene
MANUEL ARES, JR.,* MARGUERITE MANGIN, AND ALAN M. WEINER
Department ofMolecular Biophysics and Biochemistry, Yale University School ofMedicine, New Haven,
Connecticut 06510
Received 8 January 1985/Accepted 25 March 1985
We examined the structure of the promoter for the human U2 snRNA gene, a strong RNA polymerase II
transcription unit without an obvious TATA box. A set of 5' deletions was constructed and assayed for the
ability to direct initiation of U2 snRNA after microinjection into Xenopus oocytes. Sequences between positions
-295 and -218 contain an activator element which stimulates accurate initiation by 20- to 50-fold, although
as few as 62 base pairs of 5' flanking sequence are sufficient to direct the accurate initiation of U2 RNA. When
the activator was recloned in the proper orientation at either of two different upstream locations, the use of the
normal U2 start site was stimulated. Inversion of the element destroyed the stimulation of accurate U2
initiation, but initiation at aberrant upstream start sites was enhanced by the element in both orientations. A
4-base-pair deletion that destroyed the activity of the element lies within a sequence (region III) which is highly
conserved among U2 genes from different organisms. Mutations in the activator also affected the ability of the
U2 template to compete with a wild-type Ul gene in coinjection experiments. We propose that the element

  

Source: Ares Jr., Manny - Department of Molecular, Cell, and Developmental Biology, University of California at Santa Cruz

 

Collections: Biology and Medicine