Late cytoplasmic assembly intermediates are regulated in multiple layers. (Blue) 40S subunits; (green) 60S subunits; (orange) energy-consuming assembly factors; (yellow) select other assembly factors. Phosphorylation of Enp1, Rps3, and Tif6, catalyzed by Hrr25, and subsequent dephosphorylation, are required for maturation and export of the 40S and 60S subunits, respectively. Nob1-dependent cleavage at the 3′-end of 18S rRNA is regulated by the kinases Rio1 and Rio2, and the ATPase Fap7. The binding site for the bacterial homolog of Dim1, KsgA, overlaps with the binding site for IF3, indicating that Dim1-containing ribosomes cannot initiate translation. Rli1 binds to both pre-ribosomes and eIF3, indicating a link between assembling and initiating ribosomes. Release of Tif6 requires binding of the Shwachman-Diamond protein Sdo1, perhaps to recruit the GTPase Ria1, which can remove Tif6 from ribosoms in vitro. In addition, Drg1 is required to release Tif6 from pre-ribosomes. Arb1 binds to pre-ribosomes containing Tif6, but also interacts with initiation factors, linking the ribosome assembly and translation machineries.