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Acetyl-CoA is utilized in the cytosol of plant cells for the synthesis of a diverse set of phytochemicals including waxes, isoprenoids, stilbenes and flavonoids. The source of cytosolic acetyl-CoA is unclear. We identified two Arabidopsis cDNAs, which encode proteins similar to the amino and carboxy portions of human ATP-citrate lyase. Coexpression of these cDNAs in Saccharomyces cerevisiae confers ATP-citrate lyase activity, indicating that both the Arabidopsis genes are required for ACL activity. Arabidopsis ATPcitrate lyase is a heteromeric enzyme composed of two distinct subunits, ACLA (45 kD) and ACLB (65 kD). The holoprotein has a molecular weight of 500 kDa, which corresponds to a heterooctomer with an A4B4 configuration. ATP-citrate lyase activity and the ACLA and ACLB polypeptides are located in the cytosol, consistent with the lack of targeting peptides in the ACLA and ACLB sequences. In the Arabidopsis genome three genes encode for the ACLA subunit (ACLA-1, Atlgl0670; ACLA-2, Atlg60810; ACLA-3, Atlg09430) and two genes encode the ACLB subunit (ACLB-1, At3g06650; ACLB-2, At5g49460). The ACLA

ATP-citrate lyase has an essential role in cytosolic acetyl-CoA production in Arabidopsis