FIG 7.

Analysis of the effect of tethering Edc3 or Rps28a on RPS28B mRNA decay. A DNA fragment containing two MS2 coat protein binding sites was inserted into the 3′ UTR originally containing the cis-regulatory element of the RPS28B gene. The resulting RPS28B-MS2 allele and each of the individual alleles harboring Edc3 or Rps28a MS2 coat protein fusions were cointroduced into EDC3 and edc3Δ cells in DCP1/DCP2 (CFY159 and CFY161), dcp1Δ (HFY1067 and SYY2420), and dcp2Δ (CFY1016 and CFY1052) backgrounds. The steady-state levels of the transcript encoded by the RPS28B-MS2 allele in these cells were analyzed by Northern blotting, using an oligonucleotide probe complementary to the MS2 coat protein binding sites. The blots were also hybridized to a random-primed probe specific for SCR1 RNA to serve as loading controls. The relative levels of RPS28B-MS2 mRNA in the cells in different strain backgrounds were normalized to the respective EDC3 cells harboring the empty vector only. (A) Schematic representations of the RPS28B and RPS28B-MS2 alleles. (B) Schematic representations of the Edc3 and Rps28a MS2 coat protein fusions used in the experiment. (C to E) Analysis of the effects of tethering Edc3 or Rps28a on RPS28B mRNA decay in EDC3 and edc3Δ cells in DCP1 DCP2 (C), dcp1Δ (D), and dcp2Δ (E) backgrounds.