Nucleic Acids Research, Vol 25, Issue 3 466-474, Copyright © 1997 by Oxford University Press
EM Klenova, RH Nicolas, S U, AF Carne, RE Lee, VV Lobanenkov and GH Goodwin
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.
ARTICLES
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
Genetics Unit, Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK. klenovae@bioch.ox.ac.uk
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