Nucleo-cytoplasmic transport of U snRNPs: definition of a nuclear location signal in the Sm core domain that binds a transport receptor independently of the m3G cap.
We have investigated the nuclear transport of U1 and U5 snRNPs by microinjection studies in oocytes from Xenopus laevis using snRNP particles prepared by reconstitution in vitro. Competition studies with snRNPs showed that the Sm core domain of U1 snRNPs contains a nuclear location signal that acts independently of the m3G cap. The transport of U1 snRNP can be blocked by saturation with competitor U1 snRNPs or by U5 snRNPs, which indicates that the signals on the respective Sm core domains interact with the same transport receptors. Further, by using a minimal U1 snRNP particle reconstituted in vitro and containing only the Sm core RNP domain and lacking stem-loops I to III of U1 RNA, we show that this is targeted actively to the nucleus, in spite of the absence of the m3G cap. This indicates that under certain conditions the NLS in the Sm core domain not only is an essential, but may also be a sufficient condition for nuclear targeting. We propose that the RNA structure of a given snRNP particle determines at least in part whether the particle's m3G cap is required for nuclear transport or can be dispensed with.
Fischer
U
U
Institut für Molekularbiologie und Tumorforschung, Philipps-Universität Marburg, Germany.
Sumpter
V
V
Sekine
M
M
Satoh
T
T
Lührmann
R
R
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D001692:Biological Transport; D002467:Cell Nucleus / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism*; D006151:Guanosine / Q000031:analogs & derivatives; D008969:Molecular Sequence Data; D012313:RNA; D012315:RNA Caps; D017412:Ribonucleoprotein, U1 Small Nuclear / Q000378:metabolism*; D017415:Ribonucleoprotein, U5 Small Nuclear / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000378:metabolism*; D014982:Xenopus laevis
D012315:RNA Caps; D017412:Ribonucleoprotein, U1 Small Nuclear; D017415:Ribonucleoprotein, U5 Small Nuclear; D012261:Ribonucleoproteins; D006151:Guanosine; C017778:N(2),N(2),7-trimethylguanosine; D012313:RNA