Elsevier

Plant Science

Volume 116, Issue 1, 19 April 1996, Pages 47-57
Plant Science

A novel basic region/helix-loop-helix protein binds to a G-box motif CACGTG of the bean seed storage protein β-phaseolin gene

https://doi.org/10.1016/0168-9452(96)04366-XGet rights and content

Abstract

Expression of the bean seed storage protein β-phaseolin is under strict developmental control primarily at the level of transcription. A G-box motif CACGTG has been shown to be a major positive cis-acting element of the β-phaseolin gene and to be recognized by a DNA binding protein from bean seed nuclei. To understand the molecular nature of the G-box-binding protein, we screened a cDNA expression library from immature bean seed and identified a positive clone based on binding activity of the proteins expressed in Escherichia coli to an oligodeoxyribonucleotide probe. DNA sequence analysis showed that the cDNA for the phaseolin G-box-binding protein (PG1) encodes a basic region/helix-loop-helix (bHLH) domain near the carboxyl terminus. Sequence comparison with other known plant bHLH proteins such as maize RB and Antirrhinum DEL suggests that PG1 represents a new member of bHLH protein family in plants. Electrophoresis mobility-shift and DNase I footprinting assays demonstrated that PG1 protein binds preferentially to the G-box (CACGTG) and not to other two phaseolin E-box motifs (CACCTG) and CATATG). Expression of PG1 RNA was detectable in all stages of seed development as well as inflower, root, and leaf. We discuss the implications of the bHLH protein for transcriptional regulation of the β-phaseolin gene.

References (49)

  • L. Walling et al.

    Transcriptional and post-transcriptional regulation of soybean seed protein mRNA levels

  • N. Murai et al.

    Phaseolin gene from bean is expressed after transfer to sunflower via tumor-inducing plasmid vectors

    Science

    (1983)
  • C. Sengupta-Gopalan et al.

    Developmentally regulated expression of the bean β-phaseolin gene in tobacco seed

  • Z-L Chen et al.

    Functional analysis of regulatory elements in a plant embryo-specific gene

  • M.M. Bustos et al.

    Positive and negative cis-acting DNA domains are required for spatial and temporal regulation of gene expression by a seed storage protein promoter

    EMBO J.

    (1991)
  • M.D. Burow et al.

    Developmental control of the β-phaseolin gene requires positive, negative, and temporal seed-specific transcriptional regulatory elements and a negative element for stem and root expression

    Plant J.

    (1992)
  • S. Chamberland et al.

    The legumin boxes and the 3′ part of a soybean β-conglycinin promoter are involved in seed gene expression in transgenic tobacco plants

    Plant Mol. Biol.

    (1992)
  • J-M. Lelievre et al.

    5′-CA-TGCAT-3′ elements modulate the expression of glycinin genes

    Plant Physiol.

    (1992)
  • P.A. Lessard et al.

    Upstream regulatory sequences from two β-conglycinin genes

    Plant Mol. Biol.

    (1993)
  • T. Fujiwara et al.

    Tissue-specific and temporal regulation of a β-conglycinin gene: roles of the RY repeat and other cis-acting elements

    Plant Mol. Biol.

    (1994)
  • A. Brid

    The essentials of DNA methylation

    Cell

    (1992)
  • A. Vongs et al.

    Arabidopsis thaliana DNA methylation mutants

    Science

    (1993)
  • F.A. Bliss et al.

    Genetic control of phaseolin protein expression in seeds of common bean, Phaseolus vulgaris L.

    Qualitas Pluntarum Plant Foods Hum. Nutr.

    (1982)
  • J.L. Slightom et al.

    Nucleotide sequences from phaseolin cDNA clones: the major storage proteins from Phaseolus vulgaris are encoded by two unique gene families

    Nucleic Acids Res.

    (1985)
  • Cited by (0)

    Accession number U18348 at the GenBank.

    2

    Present address: Department of Botany, The Hebrew University, Institute of Life Science, 91904 Jerusalem, Israel

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