Expression data from Sar1 isoform overexpressing rat hepatoma cell lines

The endoplasmic reticulum (ER) is the site of secretory lipoprotein production and de novo cholesterol synthesis, yet little is known about how these activities are coordinated with each other, or with the activity of the COPII machinery, which transports ER cargo to the Golgi. The Sar1B component of this machinery is mutated in Chylomicron Retention Disorder, establishing that this Sar1 isoform secures delivery of dietary lipids into the circulation. We used microarrays to investigate the effect of overexpression of Sar1 isoforms and a constitutively active mutant form of Sar1B, Sar1B:H79G, on global gene expression in rat hepatoma cell line, McArdle RH7777 and identified a strong down-regulation of cholesterol biosynthetic gene mRNA expression in the Sar1B:H79G-, but not the wild-type Sar1A- or Sar1B-overexpressing cell lines. RNA was extracted from cell cultures of control McArdle RH7777, and transgenic lines overexpressing human Sar1-isoforms: one for Sar1A, two for Sar1B and two expressing a constitutively active mutant form of Sar1B, Sar1B:H79G, that cannot complete GTP hydrolysis, and hybridized on Affymetrix GeneChip Rat Genome 230 2.0 arrays.