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Molecular and Cellular Biology, October 2001, p. 6870-6881, Vol. 21, No. 20
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.20.6870-6881.2001

Widespread Use of TATA Elements in the Core Promoters for RNA Polymerases III, II, and I in Fission Yeast

Mitsuhiro Hamada,1,dagger Ying Huang,1 Todd M. Lowe,2 and Richard J. Maraia1,*

Laboratory of Molecular Growth Regulation, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, 20892-27531 and Department of Genetics, Stanford University School of Medicine, Stanford, California 94305-51202

Received 22 June 2001/Accepted 9 July 2001

In addition to directing transcription initiation, core promoters integrate input from distal regulatory elements. Except for rare exceptions, it has been generally found that eukaryotic tRNA and rRNA genes do not contain TATA promoter elements and instead use protein-protein interactions to bring the TATA-binding protein (TBP), to the core promoter. Genomewide analysis revealed TATA elements in the core promoters of tRNA and 5S rRNA (Pol III), U1 to U5 snRNA (Pol II), and 37S rRNA (Pol I) genes in Schizosaccharomyces pombe. Using tRNA-dependent suppression and other in vivo assays, as well as in vitro transcription, we demonstrated an obligatory requirement for upstream TATA elements for tRNA and 5S rRNA expression in S. pombe. The Pol III initiation factor Brf is found in complexes with TFIIIC and Pol III in S. pombe, while TBP is not, consistent with independent recruitment of TBP by TATA. Template commitment assays are consistent with this and confirm that the mechanisms of transcription complex assembly and initiation by Pol III in S. pombe differ substantially from those in other model organisms. The results were extended to large-rRNA synthesis, as mutation of the TATA element in the Pol I promoter also abolishes rRNA expression in fission yeast. A survey of other organisms' genomes reveals that a substantial number of eukaryotes may use widespread TATAs for transcription. These results indicate the presence of TATA-unified transcription systems in contemporary eukaryotes and provide insight into the residual need for TBP by all three Pols in other eukaryotes despite a lack of TATA elements in their promoters.

* Corresponding author. Mailing address: 6 Center Dr., Rm. 416, Bethesda, MD 20892-2753. Phone: (301) 402-3567. Fax: (301) 480-6863. E-mail: maraiar{at}mail.nih.gov.

dagger Present address: Department of Biochemistry, Saitama Medical School, Moroyama, Iruma-gun, Saitama, 350-0495, Japan.

Molecular and Cellular Biology, October 2001, p. 6870-6881, Vol. 21, No. 20
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.20.6870-6881.2001


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