Difference between revisions of "Part:BBa J176000"

Revision as of 20:54, 5 April 2012

hPCD

Polycomb chromodomain (PCD) from the human CBX8 protein (a.a. 1-63).
Alias: KAH01

Usage and Biology

Human PCD of CBX8 (orange) Crystal structure reported by Kaustov et al. 2011. Grey = H3 histone tail; Ball and stick = trimethylated lysine 27

Protein Data Bank ID 3I91
Mammalian expression vector required
Protein domain; requires promoter, start codon, stop codon, and polyA signal for proper expression

The Polycomb chromodomain (PCD) is an ancient protein motif that is conserved in many multicellular organisms (from plants to insects to humans). The motif appears in proteins involved in regulating tissue identity, including the Drosophila (fruit fly) Pc protein, and the vertebrate Chromobox (CBX) protein family. PCD has an aromatic pocket that specifically recognizes the unfolded "tail" of histone H3 when the histone is trimethylated at lysine 27. Early characterization of this domain was done by Fischle et al (2003) and Min et al. (2003). The 3D structures for several human homologues have been reported (Kaustove et al., 2011).

In its native context, the PCD targets gene silencing proteins to genes marked with histone methylation. The PCD domain can be fused to Biobrick proteins to recruit other protein domains, such as synthetic transcriptional activators, to sites of histone methylation in human cells (Haynes and Silver, 2011).

REFERENCES:

Fischle, W, Wang, Y, Jacobs, SA, Kim, Y, Allis, CD, Khorasanizadeh, S. (2003) Molecular basis for the discrimination of repressive methyl-lysine marks in histone H3 by Polycomb and HP1 chromodomains. Genes Dev. 17:1870-1881.
Min, J, Zhang, Y, Xu, RM. (2003) Structural basis for the specific binding of Polycomb chromodomain to histone H3 methylated at Lys 27. Genes Dev. 17:1823-1828.
Haynes, KA, Silver, PA. (2011) Synthetic reversal of epigenetic silencing. J Biol Chem. 286:27176-27182.
Kaustov, L., Ouyang, H., Amaya, M., Lemak, A., Nady, N., Duan, S., Wasney, G.A., Li, Z., Vedadi, M., Schapira, M., Min, J., Arrowsmith, C.H. (2011) Recognition and specificity determinants of the human cbx chromodomains. J Biol Chem. 286:521-529.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 7: / Line 7: @@
 ===Usage and Biology===
-[[Image:Haynes_CBX8_PDB3I91.png|thumb|250px|'''Human PCD''' (orange) Crystal structure reported by Kaustov et al. 2011. Grey = H3 histone tail; Ball and stick = trimethylated lysine 27 ]]
+[[Image:Haynes_CBX8_PDB3I91.png|thumb|250px|'''Human PCD of CBX8''' (orange) Crystal structure reported by Kaustov et al. 2011. Grey = H3 histone tail; Ball and stick = trimethylated lysine 27 ]]
 * Protein Data Bank ID 3I91
@@ Line 14: / Line 14: @@
-The Polycomb chromodomain (PCD) is an ancient protein motif that is conserved in many multicellular organisms (from plants to insects to humans). The motif appears in proteins involved in regulating tissue identity, including the Drosophila (fruit fly) Pc protein, and the vertebrate Chromobox (CBX) protein family. PCD has an aromatic pocket that specifically recognizes the unfolded "tail" of histone H3 when the histone is trimethylated at lysine 27. The crystal structure has been solved for the Drosophila melanogaster (fruit fly) variant of this peptide (Fischle et al., 2003; see also Min et al., 2003).
+The Polycomb chromodomain (PCD) is an ancient protein motif that is conserved in many multicellular organisms (from plants to insects to humans). The motif appears in proteins involved in regulating tissue identity, including the Drosophila (fruit fly) Pc protein, and the vertebrate Chromobox (CBX) protein family. PCD has an aromatic pocket that specifically recognizes the unfolded "tail" of histone H3 when the histone is trimethylated at lysine 27. Early characterization of this domain was done by Fischle et al (2003) and Min et al. (2003). The 3D structures for several human homologues have been reported (Kaustove et al., 2011).
 <br>In its native context, the PCD targets gene silencing proteins to genes marked with histone methylation. The PCD domain can be fused to Biobrick proteins to recruit other protein domains, such as synthetic transcriptional activators, to sites of histone methylation in human cells (Haynes and Silver, 2011).
@@ Line 22: / Line 22: @@
 # Fischle, W, Wang, Y, Jacobs, SA, Kim, Y, Allis, CD, Khorasanizadeh, S. (2003) Molecular basis for the discrimination of repressive methyl-lysine marks in histone H3 by Polycomb and HP1 chromodomains. Genes Dev. 17:1870-1881.
 # Min, J, Zhang, Y, Xu, RM. (2003) Structural basis for the specific binding of Polycomb chromodomain to histone H3 methylated at Lys 27. Genes Dev. 17:1823-1828.
-# Haynes, KA, Silver, PA. (2011) Synthetic reversal of epigenetic silencing. J Biol Chem. E-pub ahead of print.
+# Haynes, KA, Silver, PA. (2011) Synthetic reversal of epigenetic silencing. J Biol Chem. 286:27176-27182.
+# Kaustov, L.,  Ouyang, H.,  Amaya, M., Lemak, A., Nady, N., Duan, S.,  Wasney, G.A.,  Li, Z.,  Vedadi, M., Schapira, M.,  Min, J., Arrowsmith, C.H. (2011) Recognition and specificity determinants of the human cbx chromodomains. J Biol Chem. 286:521-529.
 <!-- -->