Biochemical research on oogenesis. Binding of tRNA to the nucleoprotein particles of Xenopus laevis previtellogenic oocytes.
In previtellogenic oocytes of Xenopus laevis, nearly all tRNA is included in nucleoprotein particles (thesaurisomes) sedimenting at 42 S. We evaluate the possibility of a tRNA exchange between the particles and the ribosomes during protein synthesis. We find that the particles take up tRNA after a very short incubation in vitro. In the absence of ATP, the particles preferentially bind charged tRNA. In the presence of ATP, more tRNA binds to the particles, and the sedimentation coefficient of the integrated tRNA is displaced to 45 S. When added to nonfractionated homogenates of oocytes together with ATP, poly(U) strongly stimulates the incorporation of radioactive phenylalanine into tRNA and protein. The labeled protein (polyphenylalanine) cosediments with the ribosomes, whereas most of phenylalanyl tRNA cosediments with the thesaurisomes. These data suggest that the thesaurisomes participate to some extent in protein synthesis. They release charged tRNA, thereby supplying the ribosomes with activated amino acids. Discharged tRNA is then taken up, reacylated, and stored in the particles until the next round of peptide bond formation. The aminoacylation and storage functions are probably carried out by two very unequal populations of particles. The main subclass of particles (42 S) binds and stores tRNA in an ATP-independent manner. A much smaller subclass of particles (45 S) is responsible for reacylation of discharged tRNA.
D000818:Animals; D005260:Female; D007700:Kinetics; D009698:Nucleoproteins / Q000378:metabolism*; D009865:Oocytes / Q000378:metabolism*; D009866:Oogenesis; D011072:Poly U / Q000378:metabolism; D011485:Protein Binding; D012343:RNA, Transfer / Q000378:metabolism*; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000302:isolation & purification; D014819:Vitellogenins / Q000378:metabolism; D014981:Xenopus
D009698:Nucleoproteins; D012346:RNA, Transfer, Amino Acyl; D012261:Ribonucleoproteins; D014819:Vitellogenins; C026464:tRNA(m)(Met), methionine-; D011072:Poly U; D012343:RNA, Transfer