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. Author manuscript; available in PMC: 2015 May 19.
Published in final edited form as: Curr Biol. 2014 Apr 24;24(10):1101–1106. doi: 10.1016/j.cub.2014.03.040

Figure 2. The short Ase1 protein isoforms are a consequence of intragenic transcription.

Figure 2

(A) Transcription of ASE1 short mRNA isoform begins at bp +756 to +766. Gel image for ASE1 gene specific PCR products after 5-RACE. Schematic of results from 5-RACE after cloning and sequencing of PCR bands (10 colonies each). (B) Full-length ASE1 mRNA is not required for expression of Ase1-SFs. ase1Δ cells containing WT ASE1 or ASE1-SF plasmid were grown to log phase at 30°C and released into selective media containing 200 mM HU. Cells were collected at the indicated time points and protein samples were prepared for western blotting to determine the expression of Ase1-FL and Ase1-SFs. (C) Translation of Ase1-SFs starts at M286 and M313. ase1Δ cells containing ASE1-13myc, ase1M286A-13myc, ase1M313A-13myc or ase1M286A-M313A-13myc plasmid were grown to log phase in selective media and released into 200 mM HU for 4 hrs. Cells were collected and protein samples were prepared for western blotting with anti-myc antibody. Pgk1 protein levels are shown as a loading control. See also Figure S2.