Mammalian Peptidylglycine alpha-Amidating Monooxygenase mRNA Expression Can Be Modulated by the La Autoantigen Export

@article{citeulike:2599131, abstract = {Peptidylglycine {alpha}-amidating monooxygenase (PAM; EC 1.14.17.3) catalyzes the COOH-terminal {alpha}-amidation of peptidylglycine substrates, yielding amidated products. We have previously reported a putative regulatory RNA binding protein (PAM mRNA-BP) that binds specifically to the 3' untranslated region (UTR) of PAM-mRNA. Here, the PAM mRNA-BP was isolated and revealed to be La protein using affinity purification onto a 3' UTR PAM RNA, followed by tandem mass spectrometry identification. We determined that the core binding sequence is approximately 15-nucleotides (nt) long and is located 471 nt downstream of the stop codon. Moreover, we identified the La autoantigen as a protein that specifically binds the 3' UTR of PAM mRNA in vivo and in vitro. Furthermore, La protein overexpression caused a nuclear retention of PAM mRNAs and resulted in the down-regulation of endogenous PAM activity. Most interestingly, the nuclear retention of PAM mRNA is lost upon expressing the La proteins that lack a conserved nuclear retention element, suggesting a direct association between PAM mRNA and La protein in vivo. Reporter assays using a chimeric mRNA that combined luciferase and the 3' UTR of PAM mRNA demonstrated a decrease of the reporter activity due to an increase in the nuclear localization of reporter mRNAs, while the deletion of the 15-nt La binding site led to their clear-cut cytoplasmic relocalization. The results suggest an important role for the La protein in the modulation of PAM expression, possibly by mechanisms that involve a nuclear retention and perhaps a processing of pre-PAM mRNA molecules. 10.1128/MCB.25.17.7505-7521.2005}, author = {Brenet, Fabienne and Dussault, Nadege and Borch, Jonas and Ferracci, Geraldine and Delfino, Christine and Roepstorff, Peter and Miquelis, Raymond and Ouafik, L'Houcine}, citeulike-article-id = {2599131}, citeulike-linkout-0 = {http://dx.doi.org/10.1128/MCB.25.17.7505-7521.2005}, citeulike-linkout-1 = {http://mcb.asm.org/cgi/content/abstract/25/17/7505}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/16107699}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=16107699}, day = {1}, doi = {10.1128/MCB.25.17.7505-7521.2005}, journal = {Mol. Cell. Biol.}, keywords = {codon, cytoplasm, la, luciferase, mrna, nucleotides, nucleus, protein, rna, translation}, month = {September}, number = {17}, pages = {7505--7521}, posted-at = {2008-03-26 16:27:08}, priority = {0}, title = {Mammalian Peptidylglycine {alpha}-Amidating Monooxygenase mRNA Expression Can Be Modulated by the La Autoantigen}, url = {http://dx.doi.org/10.1128/MCB.25.17.7505-7521.2005}, volume = {25}, year = {2005} }

TY - JOUR ID - citeulike:2599131 L3 - citeulike-article-id:2599131 N2 - Peptidylglycine {alpha}-amidating monooxygenase (PAM; EC 1.14.17.3) catalyzes the COOH-terminal {alpha}-amidation of peptidylglycine substrates, yielding amidated products. We have previously reported a putative regulatory RNA binding protein (PAM mRNA-BP) that binds specifically to the 3' untranslated region (UTR) of PAM-mRNA. Here, the PAM mRNA-BP was isolated and revealed to be La protein using affinity purification onto a 3' UTR PAM RNA, followed by tandem mass spectrometry identification. We determined that the core binding sequence is approximately 15-nucleotides (nt) long and is located 471 nt downstream of the stop codon. Moreover, we identified the La autoantigen as a protein that specifically binds the 3' UTR of PAM mRNA in vivo and in vitro. Furthermore, La protein overexpression caused a nuclear retention of PAM mRNAs and resulted in the down-regulation of endogenous PAM activity. Most interestingly, the nuclear retention of PAM mRNA is lost upon expressing the La proteins that lack a conserved nuclear retention element, suggesting a direct association between PAM mRNA and La protein in vivo. Reporter assays using a chimeric mRNA that combined luciferase and the 3' UTR of PAM mRNA demonstrated a decrease of the reporter activity due to an increase in the nuclear localization of reporter mRNAs, while the deletion of the 15-nt La binding site led to their clear-cut cytoplasmic relocalization. The results suggest an important role for the La protein in the modulation of PAM expression, possibly by mechanisms that involve a nuclear retention and perhaps a processing of pre-PAM mRNA molecules. 10.1128/MCB.25.17.7505-7521.2005 IS - 17 JF - Mol. Cell. Biol. EP - 7521 TI - Mammalian Peptidylglycine {alpha}-Amidating Monooxygenase mRNA Expression Can Be Modulated by the La Autoantigen VL - 25 SP - 7505 KW - codon KW - cytoplasm KW - la KW - luciferase KW - mrna KW - nucleotides KW - nucleus KW - protein KW - rna KW - translation AU - Brenet, Fabienne AU - Dussault, Nadege AU - Borch, Jonas AU - Ferracci, Geraldine AU - Delfino, Christine AU - Roepstorff, Peter AU - Miquelis, Raymond AU - Ouafik, L'Houcine PY - 2005/09/01/ UR - http://dx.doi.org/10.1128/MCB.25.17.7505-7521.2005 ER -