Analysis of transciption-factor binding sites in the CMV / T7 promoter portion of the CMV-SEAP vector using TranFac (CMV: 232-819 = 1-588, T7: 865-883 = 589-668)
----------------------------------------------------------------------- matrix | matrix | core | matrix | sequence name | position(str)| simil. | simil. | ----------------------------------------------------------------------- Inspecting sequence CMVT7promoter (1 - 668): V$CREBP1CJUN_01 | 139 (+) | 1.000 | 1.000 | tgACGTca V$CREB_01 | 139 (+) | 1.000 | 1.000 | TGACgtca V$CREB_01 | 139 (-) | 1.000 | 1.000 | TGACgtca V$CREBP1CJUN_01 | 139 (-) | 1.000 | 1.000 | tgACGTca V$CREBP1CJUN_01 | 192 (+) | 1.000 | 1.000 | tgACGTca V$CREB_01 | 192 (+) | 1.000 | 1.000 | TGACgtca V$CREB_01 | 192 (-) | 1.000 | 1.000 | TGACgtca V$CREBP1CJUN_01 | 192 (-) | 1.000 | 1.000 | tgACGTca V$NKX25_01 | 242 (+) | 1.000 | 1.000 | tcAAGTg V$CREBP1CJUN_01 | 275 (+) | 1.000 | 1.000 | tgACGTca V$CREB_01 | 275 (+) | 1.000 | 1.000 | TGACgtca V$CREB_01 | 275 (-) | 1.000 | 1.000 | TGACgtca V$CREBP1CJUN_01 | 275 (-) | 1.000 | 1.000 | tgACGTca V$CREL_01 | 436 (+) | 1.000 | 1.000 | ggggatTTCC V$CREBP1CJUN_01 | 461 (+) | 1.000 | 1.000 | tgACGTca V$CREB_01 | 461 (+) | 1.000 | 1.000 | TGACgtca V$CREB_01 | 461 (-) | 1.000 | 1.000 | TGACgtca V$CREBP1CJUN_01 | 461 (-) | 1.000 | 1.000 | tgACGTca
CREBP1CJUN_01 =
CRE-BP1/c-Jun = CRE-binding protein 1/c-Jun heterodimer,
Benbrook D. M., Jones N. C.: Different binding specificities and
transactivation of variant CRE's by CREB complexes. Nucleic Acids
Res. 22:1463-1469 (1994)
CREB_01 = CREB =
cAMP-responsive element binding protein,
Benbrook D. M., Jones N. C.: Different binding specificities and
transactivation of variant CRE's by CREB complexes. Nucleic Acids
Res. 22:1463-1469 (1994)
NKX25_01 = Nkx-2.5 = homeo
domain factor Nkx-2.5/Csx, tinman homolog
Chen C. Y., Schwartz R. J.: Identification of novel DNA binding
targets and regulatory domains of a murine tinman homeodomain
factor, nkx-2.5. J. Biol. Chem. 270:15628-15633 (1995) Abstract:
A murine cardiac-specific homeodomain gene named csx (Komuro, I.,
and Izumo. S.(1993) Proc. Natl. Acad. Sci. U. S. A. 90,
8145-8149) and nkx-2.5 (Lints, T. J., Parsons, L. M., Hartley,
L., Lyons, I., and Harvey, R. P.(1993) Development 119, 419-431)
was identified as a potential vertebrate homologue of Drosophila
tinman, a mesoderm determination factor required for insect heart
formation (Bodmer, R.(1993) Development 118, 719-729). Bacterial
expression of the nkx-2.5 homeodomain allowed us to identify
downstream DNA targets from a library of randomly generated
oligonucleotides. High affinity nkx-2.5 DNA binding sites,
5`-TNNAGTG-3`, represented novel binding sequences, whereas
intermediate and weaker affinity sites, 5`-C(A/T)TTAATTN-3`,
contained the typical 5`-TAAT-3` core required by most
homeodomain factors for DNA binding. We also observed that
nkx-2.5 served as a modest transcription activator in
transfection assays done in 10T1/2 fibroblasts with multimerized
binding sites linked to a luciferase reporter gene. Functional
dissection of nkx-2.5 revealed a COOH-terminal inhibitory domain
composed mainly of clusters of alanines and prolines, which
appeared to mask a potent activation domain composed of
hydrophobic and highly charged amino acids.