A homogeneous scintillation proximity assay for acetyl coenzyme A carboxylase coupled to fatty acid synthase

2006 ◽  
Vol 358 (2) ◽  
pp. 257-265 ◽  
Author(s):  
Ramakrishna Seethala ◽  
Zhengping Ma ◽  
Rajasree Golla ◽  
Dong Cheng
Keyword(s):  
Fatty Acid ◽  
Fatty Acid Synthase ◽  
Coenzyme A ◽  
Acetyl Coenzyme A ◽  
Scintillation Proximity Assay ◽  
Acetyl Coenzyme ◽  
Acetyl Coenzyme A Carboxylase

scholarly journals Inhibition of Acetyl Coenzyme A Carboxylase Activity Restores Expression of the INO1 Gene in a snf1Mutant Strain of Saccharomyces cerevisiae

2001 ◽  
Vol 21 (17) ◽  
pp. 5710-5722 ◽  
Author(s):  
Margaret K. Shirra ◽  
Jana Patton-Vogt ◽  
Andreas Ulrich ◽  
Oksana Liuta-Tehlivets ◽  
Sepp D. Kohlwein ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Fatty Acid ◽  
Fatty Acid Biosynthesis ◽  
Coenzyme A ◽  
Phospholipid Biosynthesis ◽  
Acid Biosynthesis ◽  
Acetyl Coenzyme A ◽  
Acetyl Coenzyme ◽  
Soraphen A ◽  
Acetyl Coenzyme A Carboxylase

ABSTRACT Mutations in the Saccharomyces cerevisiae SNF1 gene affect a number of cellular processes, including the expression of genes involved in carbon source utilization and phospholipid biosynthesis. To identify targets of the Snf1 kinase that modulate expression of INO1, a gene required for an early, rate-limiting step in phospholipid biosynthesis, we performed a genetic selection for suppressors of the inositol auxotrophy ofsnf1Δ strains. We identified mutations inACC1 and FAS1, two genes important for fatty acid biosynthesis in yeast; ACC1 encodes acetyl coenzyme A carboxylase (Acc1), and FAS1 encodes the β subunit of fatty acid synthase. Acc1 was shown previously to be phosphorylated and inactivated by Snf1. Here we show thatsnf1Δ strains with increased Acc1 activity exhibit decreased INO1 transcription. Strains carrying theACC1 suppressor mutation have reduced Acc1 activity in vitro and in vivo, as revealed by enzymatic assays and increased sensitivity to the Acc1-specific inhibitor soraphen A. Moreover, a reduction in Acc1 activity, caused by addition of soraphen A, provision of exogenous fatty acid, or conditional expression ofACC1, suppresses the inositol auxotrophy ofsnf1Δ strains. Together, these findings indicate that the inositol auxotrophy of snf1Δ strains arises in part from elevated Acc1 activity and that a reduction in this activity restores INO1 expression in these strains. These results reveal a Snf1-dependent connection between fatty acid production and phospholipid biosynthesis, identify Acc1 as a Snf1 target important forINO1 transcription, and suggest models in which metabolites that are generated or utilized during fatty acid biosynthesis can significantly influence gene expression in yeast.

Sign in / Sign up

Export Citation Format

Share Document