Signals and receptors for the nuclear transport of TFIIIA in Xenopus oocytes.
The transcription factor IIIA (TFIIIA) is a zinc finger protein that binds to both 5S genes and 5S ribosomal RNA. In Xenopus oocytes it is predominantly associated with 5S rRNA and retained as storage particle (7S RNP) in the cytoplasm. In this study, we have mapped the nuclear localization signal (NLS) activity in TFIIIA both in vivo and in vitro. Two independent nuclear import signals localize to the zinc finger region of TFIIIA, which is in direct contact with 5S rRNA in the context of the 7S RNP. A systematic analysis of importin alpha variants in Xenopus reveals that only importin alpha1 and importin alpha2 are expressed in a pattern similar to TFIIIA during Xenopus embryogenesis; the same two import adaptors interact specifically with TFIIIA in vitro. On the basis of these and our previous findings, we therefore propose that the massive amounts of TFIIIA which are produced in early stages of oogenesis are imported into the nucleus via interaction with importin alpha1 and alpha2. TFIIIA-induced synthesis of 5S rRNA then allows for the formation and nuclear export of the 7S RNP; the 7S RNP is retained in the cytoplasm due to NLS masking via 5S rRNA binding.
Wischnewski
Jörg
J
Abteilung Entwicklungsbiochemie, Institute für Biochemie und Molekulare Zellbiologie, Georg-August-Universität, Göttingen, Germany.
Rudt
Falko
F
Pieler
Tomas
T
D021581:Active Transport, Cell Nucleus; D000818:Animals; D002462:Cell Membrane / Q000187:drug effects; D002463:Cell Membrane Permeability / Q000187:drug effects; D002467:Cell Nucleus / Q000378:metabolism*; D004072:Digitonin / Q000494:pharmacology; D017353:Gene Deletion; D006367:HeLa Cells; D006801:Humans; D007202:Indicators and Reagents / Q000494:pharmacology; D028884:Karyopherins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008845:Microinjections; D019913:Nuclear Localization Signals / Q000737:chemistry*; D009865:Oocytes / Q000378:metabolism*; D012341:RNA, Ribosomal, 5S / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012261:Ribonucleoproteins / Q000378:metabolism; D035582:Transcription Factor TFIIIA / Q000737:chemistry / Q000378:metabolism*; D014981:Xenopus; D016335:Zinc Fingers
D007202:Indicators and Reagents; D028884:Karyopherins; D019913:Nuclear Localization Signals; D012341:RNA, Ribosomal, 5S; D011993:Recombinant Fusion Proteins; D012261:Ribonucleoproteins; D035582:Transcription Factor TFIIIA; D004072:Digitonin