La protein is associated with terminal oligopyrimidine mRNAs in actively translating polysomes.
La is an abundant, mostly nuclear, RNA-binding protein that interacts with regions rich in pyrimidines. In the nucleus it has a role in the metabolism of several small RNAs. A number of studies, however, indicate that La protein is also implicated in cytoplasmic functions such as translation. The association of La in vivo with endogenous mRNAs engaged with polysomes would support this role, but this point has never been addressed yet. Terminal oligopyrimidine (TOP) mRNAs, which code for ribosomal proteins and other components of the translational apparatus, bear a TOP stretch at the 5' end, which is necessary for the regulation of their translation. La protein can bind the TOP sequence in vitro and activates TOP mRNA translation in vivo. Here we have quantified La protein in the cytoplasm of Xenopus oocytes and embryo cells and have shown in embryo cells that it is associated with actively translating polysomes. Disruption of polysomes by EDTA treatment displaces La in messenger ribonucleoprotein complexes sedimenting at 40-60 S. The results of polysome treatment with either low concentrations of micrococcal nuclease or with high concentrations of salt indicate, respectively, that La association with polysomes is mediated by mRNA and that it is not an integral component of ribosomes. Moreover, the analysis of messenger ribonucleoprotein complexes dissociated from translating polysomes shows that La protein associates with TOP mRNAs in vivo when they are translated, in line with a positive role of La in the translation of this class of mRNAs previously observed in cultured cells.
Cardinali
Beatrice
B
Istituto di Biologia Cellulare CNR, Via Ramarini 32, 00016 Monterotondo Scalo, Italy. bcardinali@ibc.cnr.it
Carissimi
Claudia
C
Gravina
Paolo
P
Pierandrei-Amaldi
Paola
P
D000818:Animals; D001324:Autoantigens; D001483:Base Sequence; D017931:DNA Primers; D004492:Edetic Acid / Q000494:pharmacology; D004625:Embryo, Nonmammalian / Q000502:physiology; D005260:Female; D007700:Kinetics; D008969:Molecular Sequence Data; D009865:Oocytes / Q000502:physiology*; D011132:Polyribosomes / Q000378:metabolism*; D014176:Protein Biosynthesis; D038641:RNA 5' Terminal Oligopyrimidine Sequence / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012261:Ribonucleoproteins / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism; D014982:Xenopus laevis / Q000196:embryology; D000097585:SS-B Antigen
D001324:Autoantigens; D017931:DNA Primers; D004492:Edetic Acid; D012333:RNA, Messenger; D012261:Ribonucleoproteins; D014157:Transcription Factors; D000097585:SS-B Antigen