Transcription coactivators

Details Transcription coactivators Transcription coactivators

1999-06-29

2001-06-19

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

06248584

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 VP16 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 adaptors or coactivators. These transcription coactivators normally cannot 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 Epstein-Barr virus nuclear antigen 2 (EBNA 2) activates transcription of specific genes essential for B-lymphocyte transformation. EBNA 2 has an acidic activation domain which interacts with general transcription factors TFIIB, TFIIH, and TAF40. It has been shown that EBNA 2 is specifically bound to a novel nuclear protein, p100, and that p100 can coactivate gene expression mediated by the EBNA 2 acidic domain. Interestingly, p100 also appears to be essential for normal cell growth, since it has been shown that cell viability is reduced by antisense p100 RNA and restored by sense p100 RNA expression (Tong et al., (1995)
Mol. Cell Biol.
15(9):4735-4744).
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 a P100 transcription coactivator and an isolated nucleic acid fragment that is substantially similar to an isolated nucleic acid fragment encoding a P100 transcription coactivator. In addition, this invention relates to a nucleic acid fragment that is complementary to the nucleic acid fragment encoding P100.
An additional embodiment of the instant invention pertains to a polypeptide encoding all or a substantial portion of a P100 transcription coactivator.
In another embodiment, the instant invention relates to a chimeric gene encoding a P100 transcription coactivator, or to a chimeric gene that comprises a nucleic acid fragment that is complementary to a nucleic acid fragment encoding a P100 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 a P100 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 a P100 transcription coactivator in a transformed host cell comprising: a) transforming a host cell with a chimeric gene comprising a nucleic acid fragment encoding a P100 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 P100 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 a P100 transcription coactivator.
BRIEF DESCRIPTION OF THE SEQUENCE DESCRIPTIONS
The invention can be more fully understood from the following detailed description and the accompanying Sequence Listing which form a part of this application.
Table 1 lists the polypeptides that are described herein, the designation of the cDNA clones that comprise the nucleic acid fragments encoding polypeptides representing all or a substantial portion of these polypeptides, and the corresponding identifier (SEQ ID NO:) as used in the attached Sequence Listing. The sequence descriptions and Sequence Listing attached hereto comply with the rules governing nucleotide and/or amino acid sequence disclosures in patent applications as set forth in 37 C.F.R. §1.821-1.825.
TABLE 1
Transcription Coactivators
SEQ ID NO:
Protein
Clone Designation
(Nucleotide)
(Amino Acid)
P100
Contig composed of:
1
2
p0079.ctxme60r
p0119.cmtng79r
p0119.cmtnr54r
p0125.czaay64r
P100
Contig composed of:
3
4
rca1c.pk005.i19
rca1n.pk031.h24
P100
Contig composed of:
5
6
sdp3c.pk011.b23
sdp3c.pk019.h1
se1.pk0045.a4
sr1.pk0006.c7
ss11c.pk001.i10
ssm.pk0068.c2
P100
Contig composed of:
7
8
wr1.pk0030.f12
wr1.pk148.c6
The Sequence Listing contains the one letter code for nucleotide sequence characters and the three letter codes for amino acids as defined in conformity with the IUPAC-IUBMB standards described in
Nucleic Acids Research
13:3021-3030 (1985) and in the
Biochemical Journal
219 (No. 2):345-373 (1984) which are herein incorporated by reference. The symbols and format used for nucleotide and amino acid sequence data comply with the rules set forth in 37 C.F.R. §1.822.
DETAILED DESCRIPTION OF THE INVENTION
In the context of this disclosure, a number of terms shall be utilized. As used herein, a “nucleic acid fragment” is a polymer of RNA or DNA that is single- or double-stranded, optionally containing synthetic, non-natural or altered nucleotide bases. A nucleic acid fragment in the form of a polymer of DNA may be comprised of one or more segments of cDNA, genomic DNA or synthetic DNA.
As used herein, “contig” refers to a nucleotide sequence that is assembled from two or more constituent nucleotide sequences that share common or overlapping regions of sequence homology. For example, the nucleotide sequences of two or more nucleic acid fragments can be compared and aligned in order to identify common or overlapping sequences. Where common

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