Human transcription factor IIA

Details Human transcription factor IIA Human transcription factor IIA

1997-04-22

2002-02-05

McGarry, Sean (Department: 1635)

Chemistry: natural resins or derivatives; peptides or proteins;

Proteins, i.e., more than 100 amino acid residues

Blood proteins or globulins, e.g., proteoglycans, platelet...

C530S388100, C530S387300, C530S389100, C530S387900, C530S388150, C435S007100

Reexamination Certificate

active

06344543

ABSTRACT:

This invention relates to newly identified polynucleotides, polypeptides encoded by such polynucleotides, the use of such polynucleotides and polypeptides, as well as the production of such polynucleotides and polypeptides. More particularly, the polypeptide of the present invention is the small (&ggr;) subunit of human transcription factor IIA, sometimes hereinafter referred to as “small subunit”. The invention also relates to inhibiting the action of such polypeptides.
In prokaryotes, simply mixing purified RNA polymerase, a template carrying a promoter, nucleoside triphosphates, and appropriate buffer and salts is sufficient to obtain specific gene transcription in vitro beginning at the correct sites. Purified RNA polymerase from eukaryotes, however, initiates transcription very poorly and essentially at random. Accordingly, accessory factors are required for accurate initiation of transcription in eukaryotes. Some of these transcription factors are general factors required for initiation at all promoters, while others are gene-specific and are required only for certain promoters. Among the general factors is a protein called Transcription Factor IID “TFIID”, which binds to a TATA sequence, wherein T represents thymidine and A represents adenosine, in promoters. Other general factors are also involved in the assembly of a multicomponent protein complex at the promoter.
In general, transcription factors are found to contain two functional domains, one for DNA-binding and one for transcriptional activation. These functions often reside within circumscribed structural domains that retain their function when removed from their natural context. The DNA-binding domains of transcription factors fall into several structural families based on their primary amino acid sequence.
In order to identify the specific nucleotides that control gene expression, regions of the gene flanking the coding region can be sequenced. Comparisons of these sequences reveal common patterns near the 5′ and 3′ ends of different genes. These are predicted to be important for proper transcription by RNA polymerase. The most common motif is the TATA sequence around 30 bp from the transcriptional start site. Other conserved sequences have been found roughly 50 to 100 bp upstream of the transcriptional start site.
Eukaryotic transcriptional activation requires the characterization of several multiprotein complexes, referred as general transcription factors and coactivators
1,2
. The heteromeric general transcription factor TFIIA binds directly to the TATA binding protein (TBP)
3,4
and has been implicated in the process of transcriptional activation
5-8
. The &ggr; subunit of TFIIA binds weakly to the TATA binding protein, but strongly stabilized the binding of the large subunit of TFIIA (&agr; &bgr;) to TBP. Recombinant human TFIIA is functional for the transcriptional activation mediated by at least three distinct activators. Both the &agr; &bgr; and &ggr; subunits are essential for activator dependent stimulation of TFIID by binding to promoter DNA, thus facilitating the first step in pre-initiation complex formation. This demonstrates that TFIIA is an evolutionary conserved general transcription factor important for activator regulated transcription.
The interaction of TFIIA with the general transcription factor IID (TFIID) has been shown to be rate-limiting step in the transcriptional activation process
5
. TFIIA binds directly to TBP
3,4
, the DNA binding subunit of the multiprotein TFIID complex
12
. TBP associated factors (TAFs)
12-14
, which are essential for activated transcription, are also required for an activator-dependent stimulation of the TFIIA-TFIID-promoter complex
6
. While TFIIA has no known function in unregulated basal transcription
15
, it has been postulated that TFIIA plays a role in preventing inhibitors of TFIID from repressing transcription
16-19
.
A TFIIA homolog has been identified in yeast
9-10
, and the genes encoding the two subunits are essential for viability
11
. Human TFIIA consists of three polypeptides (&agr;, &bgr;, &ggr;), but the two largest subunits are derived from a single gene which shares homology to the large subunit of yeast TFIIA
7,20,21
. Both the human and yeast protein bind to the evolutionary conserved domain of TBP
22
and stimulate transcription reconstituted with TFIID, but not TBP
17,19
.
The polypeptide of the present invention has been putatively identified as the &ggr; subunit of TFIIA. This identification has been made as a result of amino acid sequence homology.
In accordance with one aspect of the present invention, there is provided a novel mature polypeptide which is the small subunit of TFIIA, as well as fragments, analogs and derivatives thereof. The polypeptide of the present invention is of human origin.
In accordance with another aspect of the present invention, there are provided polynucleotides (DNA or RNA) which encode such polypeptide.
In accordance with yet a further aspect of the present invention, there is provided a process for producing such polypeptide by recombinant techniques.
In accordance with yet a further aspect of the present invention, there is provided a process for utilizing such polypeptide, or polynucleotide encoding such polypeptide for therapeutic purposes, for example, for regulating gene-specific or global transcription and to counteract repressors of the TFIID complex.
In accordance with yet a further aspect of the present invention, there are provided antibodies against such polypeptides.
In accordance with yet another aspect of the present invention, there are provided antagonists to such polypeptides, which may be used to inhibit the action of such polypeptides, for example, to inhibit transcription of undesired cells, e.g., malignancies.
These and other aspects of the present invention should be apparent to those skilled in the art from the teachings herein.


REFERENCES:
patent: 4981790 (1991-01-01), Haseltine et al.
Copy of Geneseq/Derwent file T26459 showing sequence HUMGSO8701 in WO 95/14772 Oct. 23, 1996.
Buratowski et ,al. (1994), Cell, vol. 77, pp. 1-3.
Buratowski et ,al. (1989), Cell, vol. 56 (4), pp. 549-561.
Gould, et al. (1989), Proc. Nat'l Acad. Sci., vol. 86, pp. 1934-1938.
Matsudaira , et al. (1987), The J . Biol. Chem, vol. 262 (21), pp. 10035-10038.
Lathe, et al. (1985), J. Mol. Biol., vol. 183, pp. 1-12.
Tijan, et al. (1994), Cell, vol. 77, pp. 5-8.
Ranish, et al (1992), Science, vol. 255, pp. 1127-1129.
Cortes, et al. (1992), Molecular and Cellular Biology, vol. 12 (1) pp. 413-421.
DeJong, et al. (1995), Proc. Natl. Acad. Sci. U.S.A., vol. 92 (8), pp. 3313-3317.
Sun, et al. (1994), Genes Dev., vol. 8 (19), pp. 2336-2348.
Ozer, et al. (1994), Genes Dev., vol. 8 (19), pp. 2324-2335.
Yokomori, et al. (1994), Genes Dev., vol. 8 (19), pp. 2313-2323.
Imbalzano, et al (1994), J. Biol. Chem., vol. 269 (11), pp. 8280-8286.
Ma, et al. (1993), Genes Dev., vol. 7 (11), pp. 2246-2257.
Yokomori, et al. (1993), Genes Dev., vol. 7 (11), pp. 2235-2345.
DeJong, et al. (1993), Genes Dev., vol. 7 (11), pp. 2220-2234.
Bernstein, et al. (1994), J. Biol. Chem., vol. 269 (39), pp. 24361-24366.

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