Smad6 and uses thereof

Details Smad6 and uses thereof Smad6 and uses thereof

2001-08-07

2003-03-18

Carlson, Karen Cochrane (Department: 1653)

Drug, bio-affecting and body treating compositions

Designated organic active ingredient containing

Peptide containing doai

C530S350000

Reexamination Certificate

active

06534476

ABSTRACT:

FIELD OF THE INVENTION
This invention relates to nucleic acids and encoded polypeptides which interact with the TGF-&bgr; superfamily receptor complexes and which are a negative regulators of TGF-&bgr; superfamily signalling. The invention also relates to agents which bind the nucleic acids or polypeptides. The invention further relates to methods of using such nucleic acids and polypeptides in the treatment and/or diagnosis of disease.
BACKGROUND OF THE INVENTION
During mammalian embryogenesis and adult tissue homeostasis transforming growth factor &bgr; (TGF-&bgr;) performs pivotal tasks in intercellular communication (Roberts et al.,
Growth Factors
8:1-9, 1993). The cellular effects of this pleiotropic factor are exerted by ligand-induced hetero-oligomerization of two distantly related type I and type II serine/threonine kinase receptors, T&bgr;R-I and T&bgr;R-II, respectively (Lin and Lodish,
Trends Cell Biol.
11:972-978, 1993; Derynck,
Trends Biochem. Sci.
19-:548-553, 1994; Massague and Weis-Garcia,
Cancer Surv.
27:41-64, 1996; ten Dijke et al.,
Curr. Opin. Cell. Biol.
8:139-145, 1996). The two receptors, which both are required for signalling, act in sequence; T&bgr;R-I is a substrate for the constitutively active T&bgr;R-II kinase (Wrana et al.,
Nature
370:341-347, 1994; Weiser et al.,
EMBO J.
14:2199-2208, 1995).
TGF-&bgr; is the prototype of a large family of structurally related proteins that are involved in various biological activities (Massague, et al.,
Trends Cell Biol.
7:187-192, 1997; Roberts & Sporn, in:
Peptide growth factors and their receptors, Part I
(Sporn, M. B. and Roberts, A. B., eds) pp. 319-472, Springer-Verlag, Heidelberg (1990); Yingling et al.,
Biochim. Biophys. Acta
1242:115-136, 1995). The TGF-&bgr; “superfamily” includes activins and bone morphogenetic proteins (BMPs) that signal in a similar fashion, each employing distinct complexes of type I and type II serine/threonine kinase receptors (Lin and Lodish, 1993; Derynck, 1994; Massague and Weis-Garcia, 1996; ten Dijke et al., 1996). TGF-&bgr; related molecules act in environments where multiple signals interact and are likely to be under tight spatial and chronological regulation. For example, activin and BMP exert antagonistic effects in the development of Xenopus embryos (Graff et al.,
Cell
85:479-487, 1996). Chordin (Piccolo et al.,
Cell
86:589-598, 1996) and noggin (Zimmerman et al.,
Cell
86:599-606, 1996), for example, inhibit the ventralizing effect of BMP4 by binding specifically to the ligand. Likewise, follistatin neutralizes the activity of activin (Hemmati-Brivalou et al.,
Cell
77:283-295, 1994).
Genetic studies of TGF-&bgr;-like signalling pathways in Drosophila and
Caenorhabditis elegans
have led to the identification of mothers against dpp (Mad) (Sekelsky et al.,
Genetics
139:1347-1358, 1995) and sma (Savage et al.,
Proc. Natl. Acad. Sci. USA
93:790-794, 1996) genes, respectively. The products of these related genes perform essential functions downstream of TGF-&bgr;-like ligands acting via serine/threonine kinase receptors in these organisms (Wiersdorff et al.,
Development
122:2153:2163, 1996; Newfeld et al.,
Development
122:2099-2108, 1996; Hoodless et al.,
Cell
85:489-500, 1996). Vertebrate homologs of Mad and sma have been termed Smads (Derynck et al.,
Cell
87:173, 1996) or MADR genes (Wrana and Attisano,
Trends Genet.
12:493-496, 1996). Genetic alterations in Smad2 and Smad4/DPC4 have been found in specific tumor subsets, and thus Smads may function as tumor suppressor genes (Hahn et al.,
Science
271:350-353, 1996; Riggins et al.,
Nature Genet.
13:347-349,1996; Eppert et al.,
Cell
86:543-552, 1996). Smad proteins share two regions of high similarity, termed MH1 and MH2 domains, connected with a variable proline-rich sequence (Massague,
Cell
85:947-950, 1996; Derynck and Zhang,
Curr. Biol.
6:1226-1229, 1996). The C-terminal part of Smad2, when fused to a heterologous DNA-binding domain, was found to have transcriptional activity (Liu et al.,
Nature
381:620-623, 1996; Meersseman et al.,
Mech. Dev.
61:127-1400, 1997). The intact Smad2 protein when fused to a DNA-binding domain, was latent, but transcriptional activity was unmasked after stimulation with ligand (Liu et al., 1996).
Different Smads specify different responses using functional assays in Xenopus. Whereas Smad1 induces ventral mesoderm, a BMP-like response, Smad2 induces dorsal mesoderm, an activin/TGF-&bgr;-like response (Graff et al.,
Cell
85:479-487, 1996; Baker and Harland,
Genes
&
Dev.
10:1880-1889, 1996; Thomsen,
Development
122:2359-2366, 1996). Upon ligand stimulation Smads become phosphorylated on serine and threonine residues; BMP stimulates Smad1 phosphorylation, whereas TGF-&bgr; induces Smad2 and Smad3 phosphorylation (Hoodless et al.,
Cell
85:489-500, 1996; Liu et al., 1996; Eppert et al., 1996; Lechleider et al.,
J. Biol. Chem.
271:17617-17620, 1996; Yingling et al.,
Proc. Nat'l Aced. Sci. USA
93:8940-8944, 1996; Zhang et al.,
Nature
383:168-172, 1996; Macias-Silva et al.,
Cell
87:1215-1224, 1996; Nakao et al.,
J. Biol. Chem.
272:2896-2900, 1996).
Smad4 is a common component of TGF-&bgr;, activin and BMP signalling (Lagna et al.,
Nature
383:832-836, 1996; Zhang et al.,
Curr. Biol.
7:270-276, 1997; de Winter et al.,
Oncogene
14:1891-1900, 1997). Smad4 phosphorylation has thus far been reported only after activin stimulation of transfected cells (Lagna et al., 1996). After stimulation with TGF-&bgr; or activin Smad4 interacts with Smad2 or Smad3, and upon BMP challenge a heteromeric complex of Smad4 and Smad1 has been observed (Lagna et al., 1996). Upon ligand stimulation, Smad complexes translocate from the cytoplasm to the nucleus (Hoodless et al., 1996; Liu et al., 1996; Baker and Harland, 1996; Macias-Silva et al., 1996), where they, in combination with DNA-binding proteins, may regulate gene transcription (Chen et al.,
Nature
383:691-696, 1996).
SUMMARY OF THE INVENTION
The invention provides isolated nucleic acid molecules, unique fragments of those molecules, expression vectors containing the foregoing, and host cells transfected with those molecules. The invention also provides isolated polypeptides and agents which bind such polypeptides, including antibodies. The foregoing can be used in the diagnosis or treatment of conditions characterized by the expression of a Smad6 nucleic acid or polypeptide, or lack thereof. The invention also provides methods for identifying pharmacological agents useful in the diagnosis or treatment of such conditions. Here, we present the identification of Smad6, which inhibits phosphorylation of pathway specific Smad polypeptides including Smad2 and Smad1 and inhibits the TGF-&bgr; superfamily signalling pathway such as the TGF-&bgr; and BMP signalling pathways.
According to one aspect of the invention, an isolated nucleic acid molecule is provided. The molecule hybridizes under stringent conditions to a molecule consisting of the nucleic acid sequence of SEQ ID NO:1. The isolated nucleic acid molecule codes for a polypeptide which inhibits TGF-&bgr;, activin, or BMP signalling. The invention further embraces nucleic acid molecules that differ from the foregoing isolated nucleic acid molecules in codon sequence due to the degeneracy of the genetic code. The invention also embraces complements of the foregoing nucleic acids.
In certain embodiments, the isolated nucleic acid molecule comprises a molecule consisting of the nucleic acid sequence of SEQ ID NO:3 or consists essentially of the nucleic acid sequence of SEQ ID NO:1. Preferably, the isolated nucleic acid molecule consists of the nucleic acid sequence of SEQ ID NO:3,
According to another aspect of the invention, an isolated nucleic acid molecule is provided. The isolated nucleic acid molecule comprises a molecule consisting of a unique fragment of SEQ ID NO:3 between 12 and 1487 nucleotides in length and complements thereof, provided that the isolated nucleic acid molecule excludes sequences consisting only of SEQ ID NO:4

Affiliated with

Also associated with

Rating

Smad6 and uses thereof does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Smad6 and uses thereof, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Smad6 and uses thereof will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-3003791