Nucleic Acids Research Advance Access originally published online on December 19, 2006
Nucleic Acids Research 2007 35(2):664-677; doi:10.1093/nar/gkl1037
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Nucleic Acids Research, 2007, Vol. 35, No. 2 664-677
© 2006 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 bidirectional promoter of two genes for the mitochondrial translational apparatus in mouse is regulated by an array of CCAAT boxes interacting with the transcription factor NF-Y
1 Institute of Medical Technology & Tampere University Hospital, FI-33014 University of Tampere Finland 2 Institute of Biomedical and Life Sciences, University of Glasgow Glasgow G12 8QQ, Scotland, UK
*To whom correspondence should be addressed. Tel: +35 8335517731; Fax: +35 832157710; Email: howard.t.jacobs{at}uta.fi
Received September 12, 2006. Revised November 7, 2006. Accepted November 7, 2006.
The genes for mitoribosomal protein S12 (Mrps12) and mitochondrial seryl-tRNA ligase (Sarsm and Sars2) are oppositely transcribed from a conserved promoter region of <200 bp in both human and mouse. Using a dual reporter vector we identified an array of 4 CCAAT box elements required for efficient transcription of the two genes in cultured mouse 3T3 cells, and for enforcing directionality in favour of Mrps12. Electrophoretic mobility shift assay (EMSA) and in vivo footprinting confirmed the importance of these promoter elements as sites of protein-binding, and EMSA supershift and chromatin immunoprecipitation (ChIP) assays identified NF-Y as the key transcription factor involved, revealing a common pattern of protein–DNA interactions in all tissues tested (liver, brain, heart, kidney and 3T3 cells). The inherently bidirectional activity of NF-Y makes it an especially suitable factor to govern promoters of this class, whose expression is linked to cell proliferation.
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