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Nucleic Acids Research Advance Access originally published online on May 14, 2008
Nucleic Acids Research 2008 36(11):3716-3727; doi:10.1093/nar/gkn247
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Nucleic Acids Research, 2008, Vol. 36, No. 11 3716-3727
© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Molecular Biology

The bHLH/Per-Arnt-Sim transcription factor SIM2 regulates muscle transcript myomesin2 via a novel, non-canonical E-box sequence

Susan Woods*, Alexandra Farrall, Carl Procko and Murray L. Whitelaw

School of Molecular & Biomedical Science (Biochemistry) and the Centre for the Molecular Genetics of Development, University of Adelaide, South Australia 5005, Australia

*To whom correspondence should be addressed. Tel: +618 8303 4724; Fax: +618 8303 4362; Email: susan.woods{at}ucsf.edu

Received December 28, 2007. Revised April 14, 2008. Accepted April 16, 2008.

Despite a growing number of descriptive studies that show Single-minded 2 (Sim2) is not only essential for murine survival, but also upregulated in colon, prostate and pancreatic tumours, there is a lack of direct target genes identified for this basic helix–loop–helix/PAS transcription factor. We have performed a set of microarray experiments aimed at identifying genes that are differentially regulated by SIM2, and successfully verified that the Myomesin2 (Myom2) gene is SIM2-responsive. Although SIM2 has been reported to be a transcription repressor, we find that SIM2 induces transcription of Myom2 and activates the Myom2 promoter sequence when co-expressed with the heterodimeric partner protein, ARNT1, in human embryonic kidney cells. Truncation and mutation of the Myom2 promoter sequence, combined with chromatin immunoprecipitation studies in cells, has lead to the delineation of a non-canonical E-box sequence 5'-AACGTG-3' that is bound by SIM2/ARNT1 heterodimers. Interestingly, in immortalized human myoblasts knock down of Sim2 results in increased levels of Myom2 RNA, suggesting that SIM2 is acting as a repressor in these cells and so its activity is likely to be highly context dependent. This is the first report of a direct SIM2/ARNT1 target gene with accompanying analysis of a functional response element.


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