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Molecular and Cellular Biology, March 2007, p. 1568-1580, Vol. 27, No. 5
0270-7306/07/$08.00+0     doi:10.1128/MCB.01821-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Two G-Rich Regulatory Elements Located Adjacent to and 440 Nucleotides Downstream of the Core Poly(A) Site of the Intronless Melanocortin Receptor 1 Gene Are Critical for Efficient 3' End Processing{triangledown}

Martin Dalziel,1 Nuno Miguel Nunes,1,2 and Andre Furger1*

Genetics Unit, Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, United Kingdom,1 Department of Biology, University of Aveiro, Aveiro, Portugal2

Received 26 September 2006/ Returned for modification 10 October 2006/ Accepted 11 December 2006

Cleavage and polyadenylation is an essential processing reaction required for the maturation of pre-mRNAs into stable, export- and translation-competent mature mRNA molecules. This reaction requires the assembly of a multimeric protein complex onto a bipartite core sequence element consisting of an AAUAAA hexamer and a GU/U-rich downstream sequence element. In this study we have analyzed 3' end processing of the human melanocortin 1 receptor gene (MC1R). The MC1R gene is an intron-free transcription unit, and its poly(A) site lacks a defined U/GU-rich element. We describe two G-rich sequence elements that are critical for efficient cleavage at the MC1R poly(A) site. The first element is located 30 nucleotides downstream of the cleavage site and acts as an essential closely positioned enhancer. The second G-rich region is positioned more than 440 nucleotides downstream of the MC1R processing site and is instrumental for optimal processing efficiency. Both G-rich sequences contain clusters of heterogeneous nuclear ribonucleoprotein binding motifs and act together to enhance cleavage at the MC1R poly(A) site.

* Corresponding author. Mailing address: Genetics Unit, Department of Biochemistry, South Parks Road, Oxford OX1 3QU, United Kingdom. Phone: (44) 1865 275 321. Fax: (44) 1865 275 318. E-mail: andre.furger{at}bioch.ox.ac.uk.

{triangledown} Published ahead of print on 22 December 2006.

Molecular and Cellular Biology, March 2007, p. 1568-1580, Vol. 27, No. 5
0270-7306/07/$08.00+0     doi:10.1128/MCB.01821-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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