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Isolation of cDNAs encoding for two distinct isoforms of the TATA-Box binding proteins from common bean (Phaseolus vulgaris L.)

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Abstract

The TATA-box binding protein (TBP) is one of the 4 DNA-binding proteins that has been shown to associate with the proximal promoter region (−295) of the gene for bean seed storage protein phaseolin. The −295 promoter is essential for spatial and temporal control of the phaseolin gene expression. We designed a pair of degenerated primers based on the highly conserved sequence of the carboxyl-terminal domain of yeast TBP and used PCR to amplify the corresponding sequence from the bean cDNA. By using the amplified fragment as a probe, we screened a cDNA library derived from poly A(+) RNA from developing bean seeds and isolated 2 nearly full-length cDNA clones (813 and 826 bp long). The cDNAs encode 2 distinct isoforms of bean TBP, PV1 and PV2, each with an open reading frame of 200 amino acid residues. The 2 cDNA sequences share an 85.8% overall nucleotide sequence identity, with the coding region showing a higher degree of identity (94.4%) than the 5′- and 3′-untranslated regions (69%). The deduced amino acid sequence of the bean TBP isoforms differ in only 3 amino acid residues at positions 5, 9, and 16, all located in the amino-terminal region. The carboxyl-terminal domain of 180 amino acid residues shows a high degree (>82%) of evolutionary sequence conservation with the TBP sequences from other eukaryotic species. This domain possesses the 3 highly conserved structural motifs, namely the 2 direct repeat sequences, a central basic region rich in basic amino acid residues, and a region similar to the sigma factor of prokaryote. On the basis of this and other findings, we suggest that higher plants in general may have at least 2 copies of TBP gene, presumably resulting from the global duplication of the genome.

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Abbreviations

RACE:

rapid amplification of cDNA ends

TBP:

TATA-box binding protein

TAF:

TBP-associated factor

TLF:

TBP-like factor

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Authors and Affiliations

  1. Department of Plant Pathology and Crop Physiology, Louisiana State University and LSU Agricultural Center, 70803-1720, Baton Rouge, LA, USA

    Norimoto Murai

  2. chez Busset, route de Thollon, 74500, Lugrin, France

    Pierre Vauclare

  3. National Institute of Agrobiological Sciences, Kannondai 2-1-2, 305-8602, Tsukuba, Ibaraki, Japan

    Yasushi Kawagoe

Authors
  1. Pierre Vauclare
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  2. Yasushi Kawagoe
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  3. Norimoto Murai
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Additional information

Accession numbers AF015784 and AF015785 at the GenBank.

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Vauclare, P., Kawagoe, Y. & Murai, N. Isolation of cDNAs encoding for two distinct isoforms of the TATA-Box binding proteins from common bean (Phaseolus vulgaris L.). Plant Mol Biol Rep 21, 375–389 (2003). https://doi.org/10.1007/BF02772587

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