(A) Polyadenylation of rem1 transcripts is regulated independent of splicing. Cells harboring a deletion of the chromosomal rem1 gene (Averbeck et al. 2005) were transformed with a plasmid carrying an allele from which the intron had been precisely deleted. Polyadenylation was assayed using the same primers as in Figure 1A (bottom). (B) Read-through transcripts from the Δintron rem1 allele are present throughout a meiotic time course, but their levels are reduced in mid and late meiotic cells. Transcripts that extend downstream from the meiotic poly(A) site were detected using the same 5′ primer used to assay polyadenylation and a 3′ primer 10 nt downstream from the polyadenylation site. (C) Transcripts that extend well beyond the meiotic poly(A) site can be detected in proliferating cells harboring plasmid-borne wild-type rem1. Transcripts that extend beyond the polyadenylation site were assayed by RT-PCR as diagrammed, using a common 5′ primer and a series of 3′ primers (numbers are relative to the stop codon).
