Binding of the La autoantigen to the 5' untranslated region of a chimeric human translation elongation factor 1A reporter mRNA inhibits translation in vitro.
Human translation elongation factor 1A (EF1A) is a member of a large class of mRNAs, including ribosomal proteins and other translation elongation factors, which are coordinately translationally regulated under various conditions. Each of these mRNAs contains a terminal oligopyrimidine tract (TOP) that is required for translational control. A human growth hormone (hGH) expression construct containing the promoter region and 5' untranslated region (UTR) of EF1A linked to the hGH coding region (EF1A/hGH) was translationally repressed following rapamycin treatment in similar fashion to endogenous EF1A in human B lymphocytes. Mutation of two nucleotides in the TOP motif abolished the translational regulation. Gel mobility shift assays showed that both La protein from human B lymphocyte cytoplasmic extracts as well as purified recombinant La protein specifically bind to an in vitro-synthesized RNA containing the 5' UTR of EF1A mRNA. Moreover, extracts prepared from rapamycin-treated cells showed increased binding activity to the EF1A 5' UTR RNA, which correlates with TOP mRNA translational repression. In an in vitro translation system, recombinant La dramatically decreased the expression of EF1A/hGH construct mRNA, but not mRNAs lacking an intact TOP element. These results indicate that TOP mRNA translation may be modulated through La binding to the TOP element.
Zhu
J
J
Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT 84602, USA.
Hayakawa
A
A
Kakegawa
T
T
Kaspar
R L
RL
D020121:5' Untranslated Regions / Q000737:chemistry / Q000378:metabolism; D001324:Autoantigens / Q000235:genetics* / Q000378:metabolism; D001402:B-Lymphocytes / Q000187:drug effects / Q000378:metabolism; D002460:Cell Line; D017930:Genes, Reporter; D019382:Human Growth Hormone / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006801:Humans; D020648:Peptide Elongation Factor 1 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D014176:Protein Biosynthesis / Q000187:drug effects; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012261:Ribonucleoproteins / Q000235:genetics* / Q000378:metabolism; D020123:Sirolimus / Q000494:pharmacology; D014162:Transfection; D000097585:SS-B Antigen
D020121:5' Untranslated Regions; D001324:Autoantigens; D019382:Human Growth Hormone; D020648:Peptide Elongation Factor 1; D012333:RNA, Messenger; D011994:Recombinant Proteins; D012261:Ribonucleoproteins; D020123:Sirolimus; D000097585:SS-B Antigen