Characterization of the nuclear export adaptor protein Nmd3 in association with the 60S ribosomal subunit.
The nucleocytoplasmic shuttling protein Nmd3 is an adaptor for export of the 60S ribosomal subunit from the nucleus. Nmd3 binds to nascent 60S subunits in the nucleus and recruits the export receptor Crm1 to facilitate passage through the nuclear pore complex. In this study, we present a cryoelectron microscopy (cryo-EM) reconstruction of the 60S subunit in complex with Nmd3 from Saccharomyces cerevisiae. The density corresponding to Nmd3 is directly visible in the cryo-EM map and is attached to the regions around helices 38, 69, and 95 of the 25S ribosomal RNA (rRNA), the helix 95 region being adjacent to the protein Rpl10. We identify the intersubunit side of the large subunit as the binding site for Nmd3. rRNA protection experiments corroborate the structural data. Furthermore, Nmd3 binding to 60S subunits is blocked in 80S ribosomes, which is consistent with the assigned binding site on the subunit joining face. This cryo-EM map is a first step toward a molecular understanding of the functional role and release mechanism of Nmd3.
Sengupta
Jayati
J
Wadsworth Center, New York State Department of Health, Albany, NY 12201, USA.
Bussiere
Cyril
C
Pallesen
Jesper
J
West
Matthew
M
Johnson
Arlen W
AW
Frank
Joachim
J
D021581:Active Transport, Cell Nucleus; D001665:Binding Sites; D020285:Cryoelectron Microscopy; D009687:Nuclear Proteins / Q000378:metabolism; D011485:Protein Binding; D012335:RNA, Ribosomal / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism*; D054683:Ribosome Subunits, Large, Eukaryotic / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*
C117894:NMD3 protein, S cerevisiae; D009687:Nuclear Proteins; D012335:RNA, Ribosomal; D016601:RNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins; C065039:RNA, ribosomal, 25S