Structural determinants for post-transcriptional stabilization of lactate dehydrogenase A mRNA by the protein kinase C signal pathway.
Activation of protein kinase C (PKC) and protein kinase A (PKA) in rat C6 glioma cells increases the half-life of short-lived lactate dehydrogenase (LDH)-A mRNA about 5- and 8-fold, respectively. PKA and PKC act synergistically and prolong LDH-A mRNA half-life more than 21-fold. Similar effects were observed after transfection and transcription of a globin/lactate dehydrogenase minigene consisting of a beta-globin expression vector in which the 3'-untranslated region (UTR) of beta-globin had been replaced with the LDH-A 3'-UTR. Synergism was only obtained by transcription of minigenes containing the entire 3'-UTR and did not occur when truncated 3'-UTR fragments were analyzed. Additional mutational analyses showed that a 20-nucleotide region, named PKC-stabilizing region (PCSR), is responsible for mediating the stabilizing effect of PKC. Previous studies (Tian, D., Huang, D., Short, S., Short, M. L., and Jungmann, R. A. (1998) J. Biol. Chem. 273, 24861-24866) have demonstrated the existence of a cAMP-stabilizing region in LDH-A 3'-UTR. Sequence analysis of PCSR identified a 13-nucleotide AU-rich region that is common to both cAMP-stabilizing region and PCSR. These studies identify a specific PKC-responsive stabilizing element and indicate that interaction of PKA and PKC results in a potentiating effect on LDH-A mRNA stabilization.
Short
S
S
Department of Cellular and Molecular Biology and Cancer Center, Northwestern University Medical School, Chicago, Illinois 60611-3008, USA.
Tian
D
D
Short
M L
ML
Jungmann
R A
RA
D020413:3' Untranslated Regions / Q000235:genetics; D000818:Animals; D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D004075:Diglycerides / Q000494:pharmacology; D007527:Isoenzymes; D007770:L-Lactate Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D008957:Models, Genetic; D009154:Mutation; D010957:Plasmids; D011493:Protein Kinase C / Q000378:metabolism*; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D015398:Signal Transduction; D013997:Time Factors; D014162:Transfection; D014407:Tumor Cells, Cultured; D014529:Uridine / Q000235:genetics
D020413:3' Untranslated Regions; D004075:Diglycerides; D007527:Isoenzymes; D012333:RNA, Messenger; D011994:Recombinant Proteins; C044295:1,2-dioctanoylglycerol; D000242:Cyclic AMP; D007770:L-Lactate Dehydrogenase; D017868:Cyclic AMP-Dependent Protein Kinases; D011493:Protein Kinase C; D014529:Uridine