Expression of gp100 and CDK2 in melanoma cells is not co-regulated by a shared promoter region

Stennett, Lawrence S., Riker, Adam I., Kroll, Tara M., Chamberlin, Janine, Miki, Toru, Nickoloff, Brisan J. and Le Poole, I. Caroline (2004) Expression of gp100 and CDK2 in melanoma cells is not co-regulated by a shared promoter region. Pigment Cell Research, 17 5: 525-532. doi:10.1111/j.1600-0749.2004.00180.x

Author Stennett, Lawrence S.
Riker, Adam I.
Kroll, Tara M.
Chamberlin, Janine
Miki, Toru
Nickoloff, Brisan J.
Le Poole, I. Caroline
Title Expression of gp100 and CDK2 in melanoma cells is not co-regulated by a shared promoter region
Journal name Pigment Cell Research   Check publisher's open access policy
ISSN 0893-5785
Publication date 2004-10
Sub-type Article (original research)
DOI 10.1111/j.1600-0749.2004.00180.x
Volume 17
Issue 5
Start page 525
End page 532
Total pages 8
Place of publication Malden, MA, United States
Publisher Wiley-Blackwell
Language eng
Abstract Expression of the pigmentation-associated gene PMel17 is regulated by a 1 kB promoter region shared between the PMel17 and CDK2 genes. The encoded melanosomal glycoprotein gp100 and the cell cycle regulatory protein CDK2 are transcribed in opposite directions. Luciferase reporter constructs were generated for subregions of the promoter containing 0, 1, 2 or 3 putative binding sites for transcription factors with basic helix-loop-helix (bHLH) motifs. The potential contribution of bHLH transcription factor microphthalmia transcription factor (MITF) to promoter activity was investigated by re-introducing microphthalmia into melanoma cells lacking expression. A bi-directional reporter construct was generated to investigate potential co-regulation of gp100 and CDK2 transcription. Promoter activity was assessed in presence and absence of phorbol ester tetradecanoyl phorbol 13-acetate (TPA). FACS analysis and immunohistology served to evaluate co-regulation of gp100 and CDK2 expression at the protein level. The full-length promoter, including a consensus binding site for MITF was found to contain sequences that suppressed gp100 expression. Introduction of MITF into nonexpressing 1123 melanoma cells did not restore gp100 expression levels. A lack of coregulation for gp100 and CDK2 as suggested by immunostaining was supported by findings of dissimilar expression regulation by TPA for either gene. The current study provides insight into transcriptional regulation of the PMel17 and CDK2 genes, important to identify strategies for modulating expression of gp100 and CDK2 proteins by melanoma cells.
Keyword gp100
Cyclin dependent kinase 2
Transcription factor
Gene expression
Tumor associated antigen
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: School of Medicine Publications
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