Systematic identification of factors mediating accelerated mRNA degradation in response to changes in environmental nitrogen
Fig 5
Disrupting the Lsm1-7p/Pat1p complex and translational regulation impairs clearance of GAP1 mRNA.
A) In the background is the distribution of fit GAP1 mRNA mean expression levels for all mutants in the pool. Indicated by colored points and lines are the means for individual knockout strains, as labeled. B-E), GAP1 mRNA relative to HTA1 mRNA before and 10 minutes after a glutamine upshift, in biological replicates. Lines are a log-linear regression fit. Points are dodged horizontally for clarity, but timepoints for modeling and for drawn lines are 0 and 10 minutes exactly. Wild-type is FY4, and each estimate of the GAP1/HTA1 ratio is normalized to the average ratio measured of FY4 at t = 0 for that qPCR batch. B) xrn1Δ, ccr4Δ, pop2Δ are all defective in GAP1 mRNA clearance (p-values < 0.004). C) lsm1Δ and lsm6Δ are slowed in GAP1 mRNA clearance (p-values < 0.0132 and 0.0299, respectively). D) edc3Δ is slowed in GAP1 mRNA clearance (p-value < 10−4). scd6Δ and tif4632Δ are slowed in GAP1 mRNA clearance (p-values < 10−5) and have lower levels of expression before the upshift (p-values < 0.003). E) A deletion of 150bp 3’ of GAP1 stop codon has no significant effect, but a deletion of 100bp 5’ of the start codon has a defect in GAP1 mRNA clearance (p-value < 10−4) and lower level of expression before the upshift (p-value < 0.0015).