Xenopus U3 snoRNA GAC-Box A' and Box A Sequences Play Distinct Functional Roles in rRNA Processing

doi:10.1128/MCB.21.18.6210-6221.2001

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Molecular and Cellular Biology, September 2001, p. 6210-6221, Vol. 21, No. 18
0270-7306/01/$04.00+0 DOI: 10.1128/MCB.21.18.6210-6221.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Xenopus U3 snoRNA GAC-Box A' and Box A Sequences Play Distinct Functional Roles in rRNA Processing

Anton V. Borovjagin and Susan A. Gerbi^*

Division of Biology and Medicine, Brown University, Providence, Rhode Island 02912

Received 6 April 2001/Returned for modification 16 May 2001/Accepted 4 June 2001

Mutations in the 5' portion of Xenopus U3 snoRNA were tested for function in oocytes. The results revealed a new cleavagesite (A0) in the 3' region of vertebrate external transcribedspacer sequences. In addition, U3 mutagenesis uncoupled cleavageat sites 1 and 2, flanking the 5' and 3' ends of 18S rRNA, andgenerated novel intermediates: 19S and 18.5S pre-rRNAs. Furthermore,specific nucleotides in Xenopus U3 snoRNA that are required forcleavages in pre-rRNA were identified: box A is essential forsite A0 cleavage, the GAC-box A' region is necessary for site1 cleavage, and the 3' end of box A' and flanking nucleotidesare required for site 2 cleavage. Differences between metazoanand yeast U3 snoRNA-mediated rRNA processing are enumerated. Thedata support a model where metazoan U3 snoRNA acts as a bridgeto draw together the 5' and 3' ends of the 18S rRNA coding regionwithin pre-rRNA to coordinate theircleavage.

^* Corresponding author. Mailing address: Brown University, Division of Biology & Medicine, 69 Brown St., Providence, RI 02912.Phone: (401) 863-2359. Fax: (401) 863-2421. E-mail: Susan_Gerbi{at}Brown.edu.

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