Aminopeptidase O contains a functional nucleolar localization signal and is implicated in vascular biology.
We have identified a gene trap integration into Aminopeptidase O, the gene encoding a member of the M1 family of metalloproteases. Using the betagal reporter of the gene trap vector, we have revealed that at least some ApO isoforms are expressed predominantly in embryonic and adult blood vessels leading us to propose that ApO plays a role in vascular cell biology. The protein produced from an engineered Gfp-ApO fusion cDNA localises to the nucleolus in transfected COS7 cells. We confirm that indeed the APO protein contains a functional nucleolar localisation domain by demonstrating that GFP-APO fusion proteins that lack the predicted nucleolar localisation signal are retained in the cytoplasm. We report the existence of multiple alternatively spliced Apo isoforms that differ with respect to the presence of exons encoding important functional domains. Alternative splicing predictably produces protein products with or without the catalytic domain and/or a nucleolar localisation signal and therefore likely represents an important mechanism in regulating the biological activity of APO that has been reported to cleave one of the peptides of the renin angiotensin pathway.
Axton
Richard
R
Queen's Medical Research Institute, University of Edinburgh, 47 Little France Crescent, Edinburgh EH16 4TJ, UK.
Wallis
Julie A
JA
Taylor
Helen
H
Hanks
Mark
M
Forrester
Lesley M
LM
D017398:Alternative Splicing; D000626:Aminopeptidases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D019556:COS Cells; D002466:Cell Nucleolus / Q000378:metabolism; D002522:Chlorocebus aethiops; D004622:Embryo, Mammalian / Q000378:metabolism; D042783:Endothelial Cells / Q000201:enzymology*; D004730:Endothelium, Vascular / Q000196:embryology / Q000201:enzymology* / Q000254:growth & development; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D051379:Mice; D019913:Nuclear Localization Signals / Q000378:metabolism*; D009928:Organ Specificity; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014471:Umbilical Veins / Q000166:cytology