A role for ribosomal protein L5 in the nuclear import of 5S rRNA in Xenopus oocytes.
Prior to ribosome assembly, 5S ribosomal RNA (5S rRNA) binds to ribosomal protein L5 to form a stable ribonucleoprotein particle (5S RNP). We have analyzed the role of L5 binding in the nuclear targeting of 5S rRNA in Xenopus oocytes, and have compared the nuclear import pathway of 5S RNPs with other karyophilic molecules. Nuclear import of in vitro-generated 5S RNPs was found to be sensitive to three general inhibitors of nuclear pore complex-mediated translocation: ATP depletion, chilling, and wheat germ agglutinin. The initial rate and extent of net nuclear import was threefold greater with preassembled 5S RNPs than with 5S rRNA microinjected alone, suggesting that L5 binding is a prerequisite for nuclear accumulation. Nuclear import of 5S rRNA/5S RNPs is a facilitated process dependent on limiting factors, since nuclear import exhibited saturation kinetics. Not only was nuclear import of labeled 5S rRNA reduced in the presence of excess unlabeled 5S rRNA, but also in the presence of the synthetic karyophilic protein P(lys)-BSA. In contrast, import was not inhibited by U1 small nuclear RNA (snRNA) or U3 small nucleolar RNA (snoRNA). 5S rRNA/5S RNP nuclear import therefore appears to follow a pathway of molecular interactions similar to many karyophilic proteins, but not the methylguanosine cap-dependent U1 snRNA pathway or the cap-independent U3 snoRNA pathway.
Murdoch
K J
KJ
Department of Zoology, University of Canterbury, Christchurch, New Zealand.
Allison
L A
LA
D000818:Animals; D001692:Biological Transport / Q000502:physiology; D001798:Blood Proteins / Q000493:pharmacokinetics; D002467:Cell Nucleus / Q000737:chemistry / Q000378:metabolism; D005260:Female; D008239:Lysine / Q000493:pharmacokinetics; D008845:Microinjections; D009685:Nuclear Envelope / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism*; D009865:Oocytes / Q000378:metabolism; D010761:Phosphorus Radioisotopes; D012341:RNA, Ribosomal, 5S / Q000378:metabolism*; D017412:Ribonucleoprotein, U1 Small Nuclear / Q000378:metabolism; D017411:Ribonucleoproteins, Small Nuclear / Q000378:metabolism; D012269:Ribosomal Proteins / Q000502:physiology*; D013462:Sulfur Radioisotopes; D014982:Xenopus laevis
D001798:Blood Proteins; D009687:Nuclear Proteins; D010761:Phosphorus Radioisotopes; D012341:RNA, Ribosomal, 5S; D017412:Ribonucleoprotein, U1 Small Nuclear; D017411:Ribonucleoproteins, Small Nuclear; D012269:Ribosomal Proteins; D013462:Sulfur Radioisotopes; C029742:ribosomal protein L5; D008239:Lysine