Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or...
Reexamination Certificate
1998-11-03
2001-06-26
Hutzell, Paula (Department: 1638)
Multicellular living organisms and unmodified parts thereof and
Method of introducing a polynucleotide molecule into or...
C435S320100, C435S419000, C435S468000, C536S023600, C800S298000
Reexamination Certificate
active
06252137
ABSTRACT:
FIELD OF THE INVENTION
This invention is in the field of plant molecular biology. More specifically, this invention pertains to nucleic acid fragments encoding transcirption activator proteins involved in regulation of gene expression in seeds.
BACKGROUND OF THE INVENTION
Gene expression levels are influenced by the interactions of transcription factors with proteins that are present in general transcription complexes. Transcription factors generally have an activation domain and a DNA binding domain. In addition, other non-DNA binding proteins known as coactivators interact with transcription factors and transcription complex proteins to further stimulate transcription. The proteins PvAlf of
Phaseolus vulgaris
(Bobb et al. (1995)
Plant Journal
8:101-113), ABI3 of
Arabidopsis thaliana
(Giraudat et al. (1992)
Plant Cell
4:1251-1261), and Vp1 of maize (McCarty et al. (1991)
Cell
66:895-905) and rice (Hattori et al. (1994)
Plant Molecular Biology
24:805-810) are related proteins that are involved in the regulation of transcription. They have been called transcription factors, though it is unclear whether they actually can bind to DNA. They may bind to DNA in conjunction with another protein, or may actually be a coactivator type of regulator. In any case, these related proteins are stimulators of transcription. Due to the uncertainty in classification, we refer to these proteins as transcription activators, which could either be transcription factors or transcription coactivators.
PvAlf is expressed specifically during seed development. The highest levels of expression of PvAlf mRNA are at the time when seed storage protein expression begins. In transient assays in bean cotyledons, PvAlf positively regulated the expression of the phaseolin and phytohemagglutinin promoters, the promoters of two abundantly expressed seed storage protein genes (Bobb et al., (1995)
Plant Journal
8:101-113). The activation response was shown to be mediated through sequences that are conserved in these and other seed storage protein gene promoters, called the Ry-repeats, as well as an associated CCAC sequence (Bobb et al. (1997)
Nucleic Acids Res
. 25:641-647). The N-terminal 243 amino acids of PvAlf were shown in transient assays in bean cotyledons to function as an activation domain, when fused to the DNA binding domain of the yeast Gal4 transcription factor (Bobb et al. (1995)
Plant Journal
8:101-113).
The PvAlf-related ABI3 gene product of Arabidopsis is also involved in the regulation of seed storage protein genes. Mutants in the ABI3 gene cause a reduction or loss of expression of the 2S and 12S seed storage proteins (Nambara et al. (1994)
Plant Cell Physiol
. 35:509-513; Parcy et al. (1994)
Plant Cell
6:1567-1582). Mutants in the related Vp1 gene of maize also cause reduction in seed storage protein expression (Kriz et al (1990)
Plant Physiol
. 92:538-542). In transient assays in maize protoplasts, Vp1 activated the maize Em promoter, the promoter from a gene expressed during embryo development (McCarty et al. (1991)
Cell
66:895-905). The activation domain of Vp1 was localized to the N-terminal 121 amino acids in Gal4 fusion experiments using transient assays (McCarty et al. (1991)
Cell
66:895-905). Like PvAlf, ABI3 and Vp1 expression is specific to seed development.
To date, no PvAlf, ABI3 or VP1 homologs have been reported in
Glycine max.
SUMMARY OF THE INVENTION
The instant invention relates to isolated nucleic acid fragments encoding a plant protein involved in regulation of gene expression. More particularly, this invention concerns isolated nucleic acid fragments encoding a soybean homolog of the
Phaseolus vulgaris
PvAlf transcription activator. In addition, this invention relates to nucleic acid fragments that are complementary to nucleic acid fragments encoding the
Phaseolus vulgaris
PvAlf transcription activator.
In another embodiment, the instant invention relates to a chimeric gene that comprises a nucleic acid fragment encoding a soybean homolog of the
Phaseolus vulgaris
PvAlf transcription activator, or to a chimeric gene that comprises a nucleic acid fragment that is complementary to a nucleic acid fragment encoding a soybean homolog of the
Phaseolus vulgaris
PvAlf transcription activator, the nucleic acid fragment operably linked to suitable regulatory sequences, wherein expression of the chimeric gene results in production of levels of the encoded protein in transformed host cells that are altered (i.e., increased or decreased) relative to the levels produced in untransformed host cells.
In a further embodiment, the instant invention concerns a transformed host cell comprising in its genome a chimeric gene comprising a nucleic acid fragment encoding a soybean homolog of the
Phaseolus vulgaris
PvAlf transcription activator or a chimeric gene comprising a nucleic acid fragment that is complementary to the nucleic acid fragment encoding soybean homolog of the
Phaseolus vulgaris
PvAlf transcription activator, the chimeric gene operably linked to suitable regulatory sequences. Expression of the chimeric gene results in production of altered levels of protein encoded by the operably linked nucleic acid fragment in the transformed host cell. The transformed host cell can be of eukaryotic or prokaryotic origin, and include cells derived from higher plants and microorganisms. The invention also includes transformed plants that arise from transformed host cells of higher plants, and seeds derived from such transformed plants.
An additional embodiment of the instant invention concerns a method of altering the level of expression of a plant homolog of the
Phaseolus vulgaris
PvAlf transcription activator in a transformed host cell comprising: a) transforming a host cell with a chimeric gene comprising a nucleic acid fragment encoding a soybean homolog of the
Phaseolus vulgaris
PvAlf transcription activator or a chimeric gene that comprises a nucleic acid fragment that is complementary to the nucleic acid fragment encoding a soybean homolog of the
Phaseolus vulgaris
PvAlf transcription activator; and b) growing the transformed host cell under conditions that are suitable for expression of the chimeric gene wherein expression of the chimeric gene results in production of altered levels of the plant homolog of the
Phaseolus vulgaris
PvAlf transcription activator in the transformed host cell.
An additional embodiment of the instant invention concerns a method for obtaining a nucleic acid fragment encoding all or substantially all of an amino acid sequence encoding a platnt homolog of the
Phaseolus vulgaris
PvAlf transcription activator.
REFERENCES:
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Bobb et al., Embl Sequence Database, Nov. 10, 1995, XP002101501.
Giraudat et al. (1992) Plant Cell 4:1251-1261.
McCarty et al. (1991) Cell 66:895-905.
Hattori et al. (1994) Plant Molecular Biology 24:805-810.
Bobb et al. (1997) Nucleic Acids Rec. 25:641-647.
Nambara et al. (1994) Plant Cell Physiol. 35:509-513.
Parcy et al. (1994) Plant Cell 6:1567-1582.
Kriz et al. (1990) Plant Physiol. 92:538-542.
NCBI General Identifier No. AAA87030.
Plant J. 8(3), 331-343 (1995).
Chu et al., (1975) Sci. Sin. Peking 18:659-668.
Odell et al. (1985) Nature 313:810-812.
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Harder Patricia A.
Odell Joan T.
E. I. Du Pont de Nemours and Company
Hutzell Paula
Mehta Ashwin D.
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