Transcription coactivators

Details Transcription coactivators Transcription coactivators

1999-06-29

2001-06-26

Carlson, Karen Cochrane (Department: 1653)

Chemistry: molecular biology and microbiology

Animal cell, per se ; composition thereof; process of...

C435S252300, C435S320100, C536S023100, C530S350000

Reexamination Certificate

active

06251668

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 transcription coactivators in plants and seeds.
BACKGROUND OF THE INVENTION
In eukaryotes transcription initiation requires the action of several proteins acting in concert to initiate mRNA production. Two cis-acting regions of DNA have been identified that bind transcription initiation proteins. The first binding site located approximately 25-30 bp upstream of the transcription initiation site is termed the TATA box. The second region of DNA required for transcription initiation is the upstream activation site (UAS) or enhancer region. This region of DNA is somewhat distal from the TATA box. During transcription initiation RNA polymerase II is directed to the TATA box by general transcription factors. Transcription activators which have both a DNA binding domain and an activation domain bind to the UAS region and stimulate transcription initiation by physically interacting with the general transcription factors and RNA polymerase. Direct physical interactions have been demonstrated between activators and general transcription factors in vitro, such as between the acidic activation domain of herpes simplex virus VP 16 and TATA-binding protein (TBP), TFIIB, or TFIIH (Triezenberg et al. (1988)
Gene Dev.
2:718-729; Stringer et al. (1990)
Nature
345:783-786; Lin et al. (1991)
Nature
353:569-571; Xiao et al. (1994)
Mol. Cell. Biol.
14:7013-7024).
A third factor that is involved in the interaction is the adaptor proteins. It is thought that adaptor proteins serve to mediate the interaction between transcriptional activators and general transcription factors. Functional and physical interactions have also been demonstrated between the activators and various transcription coactivators. These transcription coactivators normally can not bind to DNA directly, however they can “bridge” the interaction between transcription activators and general transcription factors (Pugh and Tjian (1990)
Cell
61:1187-1197; Kelleher et al. (1990)
Cell
61:1209-1215; Berger et al. (1990)
Cell
61:1199-1208).
In humans LEF-1 is a general transcription factor that participates in the regulation of the T-cell receptor alpha (TCR alpha) enhancer. The function of LEF 1 is dependent, in part, on a DNA binding domain that helps to induces a sharp bend in the DNA helix, and on an activation domain that stimulates transcription only in a specific context of other enhancer-binding proteins. The ALY transcriptional activator functions in this context-dependent manner and is a novel LEF 1-interacting protein. ALY is a ubiquitously expressed, nuclear protein that specifically associates with the activation domains of LEF-1 and AML-1, which is another protein component of the TCR alpha enhancer complex. In addition, ALY can increase DNA binding by both LEF-1 and AML proteins.
Overexpression of ALY stimulates the activity of the TCR alpha enhancer complex in cells, whereas down-regulation of ALY by anti-sense oligonucleotides virtually eliminates TCR alpha enhancer activity in T cells. Thus, ALY may mediate specific transcriptional activation by facilitating the functional collaboration of multiple proteins in the TCR alpha enhancer complex (Bruhn et al., (1997)
Genes Dev
11(5):640-653).
Accordingly, the availability of nucleic acid sequences encoding all or a portion of ALY transcription coactivator proteins would facilitate studies to better understand transcription in plants and ultimately provide methods to engineer mechanisms to control transcription.
SUMMARY OF THE INVENTION
The instant invention relates to isolated nucleic acid fragments encoding transcription coactivators. Specifically, this invention concerns an isolated nucleic acid fragment encoding an ALY transcription coactivator and an isolated nucleic acid fragment that is substantially similar to an isolated nucleic acid fragment encoding an ALY transcription coactivator. In addition, this invention relates to a nucleic acid fragment that is complementary to the nucleic acid fragment encoding ALY.
An additional embodiment of the instant invention pertains to a polypeptide encoding all or a substantial portion of an ALY transcription coactivator.
In another embodiment, the instant invention relates to a chimeric gene encoding an ALY transcription coactivator, or to a chimeric gene that comprises a nucleic acid fragment that is complementary to a nucleic acid fragment encoding an ALY transcription coactivator, operably linked to suitable regulatory sequences, wherein expression of the chimeric gene results in production of levels of the encoded protein in a transformed host cell that is altered (i.e., increased or decreased) from the level produced in an untransformed host cell.
In a further embodiment, the instant invention concerns a transformed host cell comprising in its genome a chimeric gene encoding an ALY transcription coactivator, operably linked to suitable regulatory sequences. Expression of the chimeric gene results in production of altered levels of the encoded protein 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 an ALY transcription coactivator in a transformed host cell comprising: a) transforming a host cell with a chimeric gene comprising a nucleic acid fragment encoding an ALY transcription coactivator; 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 ALY in the transformed host cell.
An addition embodiment of the instant invention concerns a method for obtaining a nucleic acid fragment encoding all or a substantial portion of an amino acid sequence encoding an ALY transcription coactivator.


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Stringer et al., (1990) Nature 345:783-786.
Lin et al., (1991) Nature 353:569-571.
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Bruhn et al. (1997) Genes Dev. 11(5):640-653.
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