A pre-ribosome with a tadpole-like structure functions in ATP-dependent maturation of 60S subunits.
Analyses of isolated pre-ribosomes yielded biochemical "snapshots" of the dynamic, nascent 60S and 40S subunits during their path from the nucleolus to the cytoplasm. Here, we present the structure of a pre-60S ribosomal intermediate located in the nucleoplasm. A huge dynein-related AAA-type ATPase (Rea1) and the Rix1 complex (Rix1-Ipi1-Ipi3) are components of an extended (approximately 45 nm long) pre-60S particle. Antibody crosslinking in combination with electron microscopy revealed that the Rea1 localizes to the "tail" region and ribosomal proteins to the "head" region of the elongated "tadpole-like" structure. Furthermore, in vitro treatment with ATP induces dissociation of Rea1 from the pre-60S subunits. Rea1 and the Rix1 complex could mediate ATP-dependent remodeling of 60S subunits and subsequent export from the nucleoplasm to the cytoplasm.
Nissan
Tracy A
TA
Biochemie-Zentrum der Universität Heidelberg, Im Neuenheimer Feld 328, 69120, Germany.
Galani
Kyriaki
K
Maco
Bohumil
B
Tollervey
David
D
Aebi
Ueli
U
Hurt
Ed
E
D000255:Adenosine Triphosphate / Q000378:metabolism*; D001693:Biological Transport, Active; D002466:Cell Nucleolus / Q000378:metabolism; D003432:Cross-Linking Reagents; D003593:Cytoplasm / Q000378:metabolism; D007074:Immunoglobulin G / Q000378:metabolism; D008854:Microscopy, Electron; D012322:RNA Precursors / Q000235:genetics*; D012335:RNA, Ribosomal; D012270:Ribosomes / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*
D003432:Cross-Linking Reagents; D007074:Immunoglobulin G; D012322:RNA Precursors; D012335:RNA, Ribosomal; D029701:Saccharomyces cerevisiae Proteins; D000255:Adenosine Triphosphate