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Mol. Cell. Biol., 01 1998, 576-589, Vol 18, No. 1
E Sancho, MR Vila, L Sanchez-Pulido, JJ Lozano, R Paciucci, M Nadal, M Fox, C Harvey, B Bercovich, N Loukili, A Ciechanover, SL Lin, F Sanz, X Estivill, A Valencia and TM Thomson
By means of differential RNA display, we have isolated a cDNA corresponding
to transcripts that are down-regulated upon differentiation of the goblet
cell-like HT-29-M6 human colon carcinoma cell line. These transcripts
encode proteins originally identified as CROC-1 on the basis of their
capacity to activate transcription of c- fos. We show that these proteins
are similar in sequence, and in predicted secondary and tertiary structure,
to the ubiquitin- conjugating enzymes, also known as E2. Despite the
similarities, these proteins lack a critical cysteine residue essential for
the catalytic activity of E2 enzymes and, in vitro, they do not conjugate
or transfer ubiquitin to protein substrates. These proteins constitute a
distinct subfamily within the E2 protein family and are highly conserved in
phylogeny from yeasts to mammals. Therefore, we have designated them UEV
(ubiquitin-conjugating E2 enzyme variant) proteins, defined as proteins
similar in sequence and structure to the E2 ubiquitin- conjugating enzymes
but lacking their enzymatic activity (HW/GDB- approved gene symbol, UBE2V).
At least two human genes code for UEV proteins, and one of them, located on
chromosome 20q13.2, is expressed as at least four isoforms, generated by
alternative splicing. All human cell types analyzed expressed at least one
of these isoforms. Constitutive expression of exogenous human UEV in
HT-29-M6 cells inhibited their capacity to differentiate upon confluence
and caused both the entry of a larger proportion of cells in the division
cycle and an accumulation in G2-M. This was accompanied with a profound
inhibition of the mitotic kinase, cdk1. These results suggest that UEV
proteins are involved in the control of differentiation and could exert
their effects by altering cell cycle distribution.
Copyright © 1998, American Society for Microbiology
Role of UEV-1, an inactive variant of the E2 ubiquitin-conjugating enzymes, in in vitro differentiation and cell cycle behavior of HT-29- M6 intestinal mucosecretory cells
Departamento de Biologia Molecular, Instituto de Biologia del Cancer, IMIM-CSIC, Barcelona, Spain.
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