Cloning of comparative gene identification-58 gene in avian species and investigation of its developmental and nutritional regulation in chicken adipose tissue1

Abstract

Adipose triglyceride lipase (ATGL) is the rate-limiting enzyme of lipolysis in chicken adipose tissue. Its regulation is not fully understood. Recent studies suggest ATGL may be regulated by physical protein-protein interactions. Comparative gene identification 58 (CGI-58) has been identified as an activator of ATGL in mice. The purpose of the current study was to clone and sequence the CGI-58 gene in avian species and to investigate its regulation during development, fasting, and refeeding. Here, we report the cloning and sequencing of the complete coding sequence of CGI-58 and the deduced AA sequences for the domestic chicken, turkey, and Coturnix quail. The CGI-58 protein is a 343-AA protein in the chicken and quail, and a 344-AA protein in the turkey. Sequence comparisons with the human and mouse show that the CGI-58 gene is highly conserved among avian and mammalian species, with complete identities at the predicted lipid-binding site. Cell fractionation of chicken fat cells and stromal-vascular cells revealed that CGI-58 is expressed primarily in mature adipocytes (P < 0.01). When compared in multiple organs and tissues, avian CGI-58 is expressed predominantly in the adipose tissue (P < 0.001), similar to ATGL. To understand CGI-58 expression during adipose tissue development, its mRNA expression was measured along with ATGL and stearoyl CoA desaturase (SCD-1) mRNA, an adipogenic marker, in embryos and adults. Messenger RNA expression of CGI-58 increased (P < 0.05) immediately after hatching, concurrent with peak ATGL expression. It is interesting that CGI-58 remained somewhat increased at posthatch d 11 and 33 as SCD-1 mRNA expression increased (P < 0.05). To evaluate the response of CGI-58 to nutritional status, chickens and quail were fasted for 24 h and subsequently refed. After the fasting period, CGI-58 mRNA was induced (P < 0.05) for both chickens and quail and was returned to control levels upon refeeding. The ATGL mRNA responded similarly, increasing dramatically after fasting and quickly decreasing with refeeding. The direct relationship between CGI-58 and ATGL mRNA expression indicates a role for CGI-58 in activating ATGL-mediated lipolysis in avian species.