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 Previous Article

Molecular and Cellular Biology, November 2001, p. 7872-7882, Vol. 21, No. 22
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.22.7872-7882.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Initiator Recognition in a Primitive Eukaryote: IBP39, an Initiator-Binding Protein from Trichomonas vaginalis

David R. Liston,1,dagger Audrey O. T. Lau,1 Diana Ortiz,1 Stephen T. Smale,1,2,3 and Patricia J. Johnson1,2,*

Department of Microbiology, Immunology, and Molecular Genetics,1 Molecular Biology Institute,2 and Howard Hughes Medical Institute,3 University of California, Los Angeles School of Medicine, Los Angeles, California 90095-1489

Received 7 June 2001/Returned for modification 4 August 2001/Accepted 17 August 2001

While considerable progress has been made in understanding the mechanisms of transcription in higher eukaryotes, transcription in single-celled, primitive eukaryotes remains poorly understood. Promoters of protein-encoding genes in the parasitic protist Trichomonas vaginalis, which represents one of the deepest-branching eukaryotic lineages, have a bipartite structure with gene-specific regulatory elements and a conserved core promoter encompassing the transcription start site. Core promoters in T. vaginalis appear to consist solely of a highly conserved initiator (Inr) element that is both a structural and a functional homologue of its metazoan counterpart. Using DNA affinity chromatography, we have isolated an Inr-binding protein from T. vaginalis. Cloning of the gene encoding the Inr binding protein identified a novel 39-kDa protein (IBP39). We show that IBP39 binds to both double and single Inr motifs found in T. vaginalis genes and that binding requires the conserved nucleotides necessary for Inr function in vivo. Analyses of the cloned IBP39 gene revealed no homology at the protein sequence level with identified proteins in other organisms or the presence of known DNA-binding domains. The relationship between IBP39 and Inr-binding proteins in metazoa presents interesting evolutionary questions.

* Corresponding author. Mailing address: Department of Microbiology, Immunology, and Molecular Genetics, UCLA School of Medicine, 405 Hilgard Ave., 1602 Molecular Sciences Bldg., Los Angeles, CA 90095-1489. Phone: (310) 825-4870. Fax: (310) 206-5231. E-mail: johnsonp{at}ucla.edu.

dagger Present address: Section of Microbial Pathogenesis, Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, CT 06536-0812.

Molecular and Cellular Biology, November 2001, p. 7872-7882, Vol. 21, No. 22
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.22.7872-7882.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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