Endothelin-converting enzyme-1 (ECE-1) is post-transcriptionally regulated by alternative polyadenylation.
Endothelin-converting enzyme-1 (ECE-1) is the enzyme predominantly responsible for producing active endothelin-1 (ET-1), a mitogenic peptide implicated in the aetiology of a number of diseases, including cancer. Elevated levels of ECE-1 have been observed in a range of malignancies, with high expression conferring poor prognosis and aiding the acquisition of androgen independence in prostate cancer. The mechanisms regulating the expression of ECE-1 in cancer cells are poorly understood, hampering the development of novel therapies targeting the endothelin axis. Here we provide evidence that the expression of ECE-1 is markedly inhibited by its 3'UTR, and that alternative polyadenylation (APA) results in the production of ECE-1 transcripts with truncated 3'UTRs which promote elevated protein expression. Abolition of the ECE-1 APA sites reduced protein expression from a reporter vector in prostate cancer cells, suggesting these sites are functional. This is the first study to identify ECE-1 as a target for APA, a regulatory mechanism aberrantly activated in cancer cells, and provides novel information about the mechanisms leading to ECE-1 overexpression in malignant cells.
Institute of Molecular and Cellular Biology, University of Leeds, Leeds, United Kindom.
Alison R
AR
Whyteside
Institute of Molecular and Cellular Biology, University of Leeds, Leeds, United Kindom.
Anthony J
AJ
Turner
Integrated Biosciences, School of Clinical Dentistry, University of Sheffield, Sheffield, United Kingdom ; CR-UK/YCR Sheffield Cancer Centre, University of Sheffield, Sheffield, United Kingdom.
Daniel W
DW
Lambert
D020413:3' Untranslated Regions / Q000235:genetics; D017398:Alternative Splicing; D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D016466:CHO Cells; D002460:Cell Line; D045744:Cell Line, Tumor; D006224:Cricetinae; D003412:Cricetulus; D000072396:Endothelin-Converting Enzymes; D006801:Humans; D015151:Immunoblotting; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008297:Male; D008666:Metalloendopeptidases / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D011061:Poly A / Q000235:genetics; D026723:Polyadenylation / Q000235:genetics*; D012323:RNA Processing, Post-Transcriptional; D022821:RNA Splice Sites / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D020413:3' Untranslated Regions; D022821:RNA Splice Sites; D011061:Poly A; D008156:Luciferases; D016282:Aspartic Acid Endopeptidases; D008666:Metalloendopeptidases; C000606923:ECE1 protein, human; D000072396:Endothelin-Converting Enzymes