The sup8 tRNALeu gene of Schizosaccharomyces pombe has an unusual intervening sequence and reduced pairing in the anticodon stem |
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| Authors: | M Sumner-Smith H Hottinger I Willis T L Koch R Arentzen D S?ll |
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| Institution: | (1) Department of Molecular Biophysiscs and Biochemistry, Yale University, 06511 New Haven, CT, USA;(2) Human Genetics, Yale University, 06511 New Haven, CT, USA;(3) Central Research and Development Department, E.I. du Pont de Nemours & Co., Experimental Station, 19898 Wilmington, DE, USA;(4) Present address: Allelix, Inc., 6850 Goreway Drive, L4V 1P1 Mississauga, Ontario, Canada;(5) Present address: Research Division, Nestle Products Technical Assistance Co. Ltd., Case Postale 88, CH-1814 La Tour de Peilz, Switzerland |
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| Abstract: | Summary We have cloned and sequenced the wild-type and suppressor alleles of the S. pombe sup8 tRNA gene. The wild-type allele has a leucine UAA anticodon and the suppressor (sup8-e) carries the opal suppressor anticodon UCA. The gene has a 16 base pair intervening sequence that, in the RNA, is predicted to form a secondary structure which involves base pairing to the 5 , rather than the usual 3 side of the 5 splice site. When incubated in Saccharomyces cerevisiae cell-free extracts both alleles are efficiently transcribed, the 5 leader and 3 trailer sequences are removed and CCA is added to the 3 processed end; however, the intervening sequence is not excised. This finding implies that the structural requirements of the splicing endonucleases in the two yeasts have diverged. No other tRNA genes with related sequences were detected in S. pombe DNA by hybridization, suggesting that other UUA isoacceptors may be structurally dissimilar to sup8 or that the UUA codon may be decoded by a UUG leucine isoacceptor. |
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