Nucleotide Analogs - Jena Bioscience

Human p300 and CBP (CREB binding protein) are
highly related transcriptional coactivators. Both proteins
have been identifi ed through protein interaction
assays. In addition to interacting with variety of cellular
factors and onco-proteins, loss of the wild type CBP
alleles in isolated tumors suggests that CBP/p300
might serve as tumor suppressors. The C-terminus
of p300 contains a Histone Acetyltransferase (HAT)
domain expanded from residues 1195 to 1673, an
activation domain (residues 1763-1812) that interacts
with many transcription factors and an SRC-1 domain
further down stream of the activation domain. p300 is
able to acetylate histones and many other transcription
factors, including p53, E2F, TFIIE, and TFIIF etc.
implicating diverse roles of p300 in gene regulation.
The C-terminus of p300 (residues 1135-2414)
was expressed in baculovirus system and purifi ed
by an affi nity column in combination with FPLC
chromatography.
Recombinant p300 (C-terminus) can be used 1)
for protein-protein interaction assay; 2) for in vitro
transcription assay; 3) for in vitro acetylation assay;
and 4) for cell growth assay.
Purifi ed protein is greater than 95% homogeneous
and contains no detectable proteases, DNase, and
RNase activity.
Unit defi nition:
50 units (ng) are required for reconstituted transcription
assay and 100 units are required for a protein-protein
interaction assay.
Purity: > 95% by SDS-PAGE.
Store: -80 °C
Selected references:
Stein et al. (1990) Analysis of E1A-mediated growth regulation
functions: binding of the 300-kilodalton cellular product
correlates with E1A enhancer repression function and DNA
synthesis-inducing activity. J. Virol. 64:4421.
Chrivia et al. (1993) Phosphorylated CREB binds specifi cally to
the nuclear protein CBP. Nature 365:855.
Eckner et al. (1994) Molecular cloning and functional analysis of
the adenovirus E1A-associated 300-kD protein (p300) reveals
a protein with properties of a transcriptional adaptor. Genes &
Dev. 8:869.
Ogryzko et al. (1996) The transcriptional coactivators p300 and
CBP are histone acetyltransferases. Cell 87:953.
Kung et al. (2000) Gene dose-dependent control of
hematopoiesis and hematologic tumor suppression by CBP.
Genes & Dev. 14:272.
Imhof et al. (1997) Acetylation of general transcription factors
by histone acetyltransferases. Curr. Biol. 7:689.
Gu et al. (1997) Activation of p53 sequence-specifi c DNA
binding by acetylation of the p53 C-terminal domain. Cell 90:
595.
Martinez-Balbas et al. (2000) Regulation of E2F1 activity by
acetylation. EMBO J. 19:662.
PC4
(Positive Cofactor 4,
Transcriptional Coactivator,
wild type)
human, Recombinant, E. coli
Cat. No. Amount Price (€)
PR-725 10 µg 350,--
Liquid. Supplied in 20 mM Tris-HCl, pH 7.9, 100 mM
KCl, 0.2 mM EDTA, 1 mM DTT, 20% glycerol.
The human PC4 is a non-TAF transcription coactivator
that mediates activator-dependent transcription
by RNA polymerase II in vitro through most tested
activators. The function of PC4 is apparently through
interactions with transcriptional activators and the
basal transcription machinery. It is negatively regulated
by casein kinase II phosphorylation both in vivo and
in vitro. PC4 strongly binds single stranded DNA
and regulates HSSB (RPA)-dependent SV40 DNA
replication. Recent studies indicated that PC4 can be
acetylated by several histone acetyltransferase.
Recombinant PC4 protein (wild type, 127 amino
acids) is isolated from an E. coli strain that carries
the coding sequence of human PC4 under the
control of T7 promoter and purifi ed by conventional
chromatography.
Recombinant PC4 has been utilized for in vitro
function studies, including transcription, DNA
replication, in vitro phosphorylation, gel mobility shift
assay, protein-protein interactions, and as a substrate
for in vitro acetylation.
Protein is greater than 95% homogeneous and
contains no detectable protease, DNase, and RNase
activity.
Unit defi nition:
1 unit equals 1 ng of purifi ed protein. 1 unit is suffi cient
for a gel mobility shift assay in a 20 µl reaction;
20 units are suffi cient for reconstituted transcription
assay and 100 units are suffi cient for a protein-protein
interaction assay.
Purity: > 95% by SDS-PAGE.
Store: -80 °C
Selected references:
Ge H. and Roeder R.G. (1994) Purifi cation, cloning, and
characterization of a human coactivator, PC4, that mediates
transcriptional activation of class II genes. Cell 78:513.
Ge et al. (1994) Phosphorylation negatively regulates the
function of coactivator PC4. Proc. Natl. Acad. Sci. USA 91:
12691.
Kretzschmar et al. (1994) A novel mediator of class II
gene transcription with homology to viral immediate-early
transcriptional regulators. Cell 78:525.
Ge et al. (1996) Activator-dependent transcription by
mammalian RNA polymerase II: in vitro reconstitution with
general transcription factors and cofactors. Methods Enzymol.
274:57.
Ballard et al. (1988) Identifi cation of a novel 9-kDa polypeptide
from nuclear extracts. DNA binding properties, primary
structure, and in vitro expression. J. Biol. Chem. 263:8450.
Pan et al. (1996) Transcription-positive cofactor 4 forms
complexes with HSSB (RPA) on single-stranded DNA and
infl uences HSSB-dependent enzymatic synthesis of simian
virus 40 DNA. J. Biol. Chem. 271:22111.
Kumar et al. (2001) p300-mediated acetylation of human
transcriptional coactivator PC4 is inhibited by phosphorylation.
J. Biol. Chem. 276:16804.
PC4-mt
(Positive Cofactor 4,
Transcriptional Coactivator,
F77P mutant)
human, Recombinant, E. coli
Cat. No. Amount Price (€)
PR-726 10 µg 350,--
Liquid. Supplied in 20 mM Tris-HCl, pH 7.9, 100 mM
KCl, 0.2 mM EDTA, 1 mM DTT, 20% glycerol.
The human PC4 is a non-TAF transcription coactivator
that mediates activator-dependent transcription
by RNA Polymerase II in vitro through most tested
activators. The function of PC4 is through interactions
with transcriptional activators and the basal
transcription machinery. It is negatively regulated by
Casein Kinase II phosphorylation both in vivo and in
vitro. PC4 strongly binds single stranded DNA and the
region essential for the single stranded DNA binding
activity was mapped around residue 77. A single
amino acid change at position 77 (F to P) abolishes
both ds and ssDNA binding activity.
Recombinant PC4 protein (mutant F77P, 127 amino
acids) is isolated from an E. coli strain that carries
the coding sequence of human PC4 under the
control of T7 promoter and purifi ed by conventional
chromatography.
Recombinant PC4 has been utilized for in vitro
function studies, including transcription, DNA
replication, in vitro phosphorylation, gel mobility shift
assay, protein-protein interactions, and as a substrate
for in vitro acetylation.
Protein is greater than 95% homogeneous and
contains no detectable protease, DNase, and RNase
activity.
Unit defi nition:
1 unit equals 1 ng of purifi ed protein. 1 unit is suffi cient
for a gel mobility shift assay in a 20 µl reaction;
20 units are suffi cient for reconstituted transcription
assay and 100 units are suffi cient for a protein-protein
interaction assay.
Purity: > 95% by SDS-PAGE.
Store: -80 °C
Transcription Factors
Selected references:
Ge H. and Roeder R.G. (1994) Purifi cation, cloning, and
characterization of a human coactivator, PC4, that mediates
transcriptional activation of class II genes. Cell 78:513.
Ge et al. (1994) Phosphorylation negatively regulates the
function of coactivator PC4. Proc. Natl. Acad. Sci. USA 91:
12691.
Kretzschmar et al. (1994) A novel mediator of class II
gene transcription with homology to viral immediate-early
transcriptional regulators. Cell 78:525.
Wu et al. (1998) Properties of PC4 and an RNA polymerase II
complex in directing activated and basal transcription in vitro. J.
Biol. Chem. 273:12492.
Brandsen et al. (1997) C-terminal domain of transcription
cofactor PC4 reveals dimeric ssDNA binding site. Nat. Struct.
Biol. 4:900.
Werten et al. (1998) Interaction of PC4 with melted DNA inhibits
transcription. EMBO J. 17:5103.
PC4-mt
(Positive Cofactor 4,
Transcriptional Coactivator,
serine mutations)
human, Recombinant, E. coli
Cat. No. Amount Price (€)
PR-727 10 µg 350,--
Liquid. Supplied in 20 mM Tris-HCl, pH 7.9, 100 mM
KCl, 0.2 mM EDTA, 1 mM DTT, 20% glycerol.
The human PC4 is a non-TAF transcription coactivator
that mediates activator-dependent transcription
by RNA Polymerase II in vitro through most tested
activators. Phosphorylation negatively regulates the
function of coactivator PC4. Mutational and mass
spectrometric studies suggest that the in vivo and
in vitro hyperphosphorylation of PC4 is mediated by
Casein Kinase II and restricted to the the N-terminal
serine–rich region. Phosphorylation of PC4 by Casein
Kinase II inhibits the p300-mediated acetylation. The
wild-type PC4 but not the kinase inhibitory-defi cient
mutant of PC4 (serine mutations) represses specifi c
transcription in vivo.
Recombinant PC4 protein (serine mutations,
127 amino acids) is isolated from an E. coli strain that
carries the coding sequence of human PC4 under the
control of T7 promoter and purifi ed by conventional
chromatography.
Recombinant PC4 has been utilized for in vitro
function studies, including transcription, DNA
replication, in vitro phosphorylation, gel mobility shift
assay, protein-protein interactions, and as a substrate
for in vitro acetylation.
Protein is greater than 95% homogeneous and
contains no detectable protease, DNase, and RNase
activity.
Unit defi nition:
1 unit equals 1 ng of purifi ed protein. 1 unit is suffi cient
for a gel mobility shift assay in a 20 µl reaction;
20 units are suffi cient for reconstituted transcription
assay and 100 units are suffi cient for a protein-protein
interaction assay.
Purity: > 95% by SDS-PAGE.
Store: -80 °C
Selected references:
Ge H. and Roeder R.G. (1994) Purifi cation, cloning, and
characterization of a human coactivator, PC4, that mediates
transcriptional activation of class II genes. Cell 78:513.
Kretzschmar et al. (1994) A novel mediator of class II
gene transcription with homology to viral immediate-early
transcriptional regulators. Cell 78:525.
Ge et al. (1994) Phosphorylation negatively regulates the
function of coactivator PC4. Proc. Natl. Acad. Sci. USA 91:
12691.
Kumar et al. (2001) p300-mediated acetylation of human
transcriptional coactivator PC4 is inhibited by phosphorylation.
J. Biol. Chem. 276:16804.
Schang et al. (2000) Human PC4 is a substrate-specifi c
inhibitor of RNA polymerase II phosphorylation. J. Biol. Chem.
275:6071.
Recombinant Proteins
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