Molecular weight abnormalities of the CTCF transcription factor: CTCF migrates aberrantly in SDS-PAGE and the size of the expressed protein is affected by the UTRs and sequences within the coding region of the CTCF gene.
CTCF belongs to the Zn finger transcription factors family and binds to the promoter region of c-myc. CTCF is highly conserved between species, ubiquitous and localised in nuclei. The endogenous CTCF migrates as a 130 kDa (CTCF-130) protein on SDS-PAGE, however, the open reading frame (ORF) of the CTCF cDNA encodes only a 82 kDa protein (CTCF-82). In the present study we investigate this phenomenon and show with mass-spectra analysis that this occurs due to aberrant mobility of the CTCF protein. Another paradox is that our original cDNA, composed of the ORF and 3'-untranslated region (3'-UTR), produces a protein with the apparent molecular weight of 70 kDa (CTCF-70). This paradox has been found to be an effect of the UTRs and sequences within the coding region of the CTCF gene resulting in C-terminal truncation of CTCF-130. The potential attenuator has been identified and point-mutated. This restored the electrophoretic mobility of the CTCF protein to 130 kDa. CTCF-70, the aberrantly migrating CTCF N-terminus per se, is also detected in some cell types and therefore may have some biological implications. In particular, CTCF-70 interferes with CTCF-130 normal function, enhancing transactivation induced by CTCF-130 in COS6 cells. The mechanism of CTCF-70 action and other possible functions of CTCF-70 are discussed.
Klenova
E M
EM
Genetics Unit, Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK. klenovae@bioch.ox.ac.uk
Nicolas
R H
RH
U
S
S
Carne
A F
AF
Lee
R E
RE
Lobanenkov
V V
VV
Goodwin
G H
GH
D000818:Animals; D019556:COS Cells; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D008970:Molecular Weight; D016366:Open Reading Frames; D017354:Point Mutation; D014176:Protein Biosynthesis; D014157:Transcription Factors / Q000737:chemistry* / Q000235:genetics*; D015533:Transcriptional Activation
D018076:DNA, Complementary; D014157:Transcription Factors