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Nucleic Acids Research, 1992, Vol. 20, No. 18 4881-4887
© 1992

MOLECULAR BIOLOGY

An ATF/CREB binding motif is required for aberrant constitutive expression of the MHC class II DR{alpha} promoter and activation by SV4O T-antigen

P.M. Cox+ and C.R. Goding*

Eukaryotic Transcription Laboratory, Marie Curie Research Institute, The Chart Oxted, Surrey RH8 OTL, UK

*To whom correspondence should be addressed

Received May 27, 1992. Revised August 14, 1992. Accepted August 14, 1992.

Constitutive expression of major histocompatibility complex class II (MHC II) antigens normally occurs in B-lymphocytes and antigen presenting cells of the monocyte/macrophage lineage. However, many malignant tumours and transformed cells express these proteins aberrantly. We demonstrate here that the MHC II DRa promoter is constitutlvely active both In the SV4O large T antigen transformed cell line, COS, and in CV1 cells from which they are derived. As an approach to understanding the molecular mechanisms underlying aberrant DRcr expression we have examined the cis and trans-acting requirements for DRa transcription in these cell types. Electrophoretic mobility shift assays showed that the region immediately 3' to the X-box was bound by a member of the ATF/CREB family of transcription factors. Using deletions and point mutations in the DRa promoter we demonstrate that, in contrast to B-cells, the octamer motif and conserved X- and V-boxes make only a minor contribution to promoter function while single point mutations in the ATF/CREB motif reduced transcription up to 20-fold. In addition, we show that the DRcr promoter is activated by SV4O large T-antigen and that activation requires an intact ATF/CREB motif. Similar data were obtained using B16 melanoma cells . These results suggest that the ATF/CREB motif may be a target for transcription deregulation in several transformed cell types.

* Present address: Department of Histopathology, Royal Surrey County Hospital, Egerton Road, Guildford, Surrey GU2 lxx, UK


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