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. Author manuscript; available in PMC: 2012 Jan 28.
Published in final edited form as: J Mol Biol. 2010 Nov 25;405(4):877–891. doi: 10.1016/j.jmb.2010.11.035

Figure 4.

Figure 4

BLM expression does not suppress sgs1Δ defects and BLM overexpression is detrimental to yeast cells. (A) Expression of myc-epitope tagged Sgs1 (lane 1) and BLM (lane 2) from the native chromosomal SGS1 locus or galactose-inducible overexpression of myc-epitope tagged Sgs1 (lane 4) and BLM (lane 6) in yeast cells grown in YP supplemented with 1% sucrose and 1% galactose (to induce expression, lanes 4 and 6) or without galactose (lanes 3 and 5). Both BLM and Sgs1 show signs of degradation upon overexpression (lanes 4 and 6) whereas expression from the native SGS1 promoter is stable (lanes 1 and 2). Molecular weights (MW) are indicated in kDa on the left. (B) Cells expressing BLM from the SGS1 promoter on chromosome VIII are as sensitive to HU as cells lacking Sgs1 (Δsgs1). Replacement of the natural SGS1 promoter with a galactose-inducible GAL1 promoter induces BLM overexpression and leads to increased HU sensitivity. Ten-fold dilutions of cells were spotted on media containing 1% sucrose and 1% galactose (to induce BLM overexpression) with and without 100 mM HU.