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Nuclear cap binding protein complex (CBC) is a heterodimer of a small subunit (Cbc2 in yeast) that binds the m 7 G cap and a large subunit (Sto1 in yeast) that interacts with karyopherins. In order to probe the role of cap recognition in yeast CBC function, we introduced alanine mutations (Y24A, F91A, D120A, D122A, R129A, and R133A) and N-terminal deletions (NΔ21 and NΔ42) in the cap-binding pocket of Cbc2. These lesions had no effect on vegetative growth, but they ameliorated the cold-sensitivity of  tgs1 Δ cells that lack trimethylguanosine caps (a phenotype attributed to ectopic association of CBC with the m 7 G cap of the normally TMG-capped U1 snRNA), thereby attesting to their impact on cap binding in vivo. Further studies of the Cbc2-Y24A variant revealed synthetic lethality or sickness with null mutations of proteins involved in early steps of spliceosome assembly (Nam8, Mud1, Swt21, Mud2, Ist3, and Brr1) and with otherwise benign mutations of Msl5, the essential branchpoint binding protein. Whereas the effects of weakening CBC–cap interactions are buffered by other actors in the splicing pathway during mitotic growth, the  NΔ42  allele causes a severe impediment to yeast sporulation and meiosis. RNA analysis revealed a selective defect in the splicing of  MER3  and  SAE3  transcripts in  cbc2-NΔ42  diploids during attempted sporulation. An intronless  MER3  cDNA fully restored sporulation and spore viability in the  cbc2-NΔ42  strain, signifying that  MER3  splicing is a limiting transaction. These studies reveal a new level of splicing control during meiosis that is governed by nuclear CBC.

Genetic interactions of hypomorphic mutations in the m<sup>7</sup>G cap-binding pocket of yeast nuclear cap binding complex: An essential role for Cbc2 in meiosis via splicing of <i>MER3</i> pre-mRNA

Genetic interactions of hypomorphic mutations in the m7G cap-binding pocket of yeast nuclear cap binding complex: An essential role for Cbc2 in meiosis via splicing of MER3 pre-mRNA

Genetic interactions of hypomorphic mutations in the m⁷G cap-binding pocket of yeast nuclear cap binding complex: An essential role for Cbc2 in meiosis via splicing of MER3 pre-mRNA