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HubMed - Alternative Health
Sodium-coupled Neutral Amino Acid Transporter 4 Functions as a Regulator of Protein Synthesis during Liver Development.
Hepatol Res. 2013 Jan 14;
Kondou H, Kawai M, Tachikawa K, Kimoto A, Yamagata M, Koinuma T, Yamazaki M, Nakayama M, Mushiake S, Ozono K, Michigami T
AIM: The molecular mechanisms by which hepatocyte nuclear factor (HNF) 4α regulates fetal liver development have not been fully elucidated. We screened the downstream molecules of HNF4α during liver development, and identified sodium-coupled neutral amino acid transporter (SNAT) 4. The aim of this study is to investigate the regulation of SNAT4 by HNF4α and to clarify its roles in differentiating hepatocytes. METHODS: HNF4αwas over-expressed in cultured liver buds using adenovirus, and suppression subtractive hybridization screening was performed. Temporal and spatial expression of SNAT4 during liver development was investigated. Regulation of SNAT4 by HNF4αwas examined by promoter analyses and electrophoretic mobility-shift assays (EMSA). Metabolic labeling and Western blotting were carried out using primary hepatoblasts with SNAT4 over-expression. RESULTS: The expression of Slc38a4 encoding SNAT4 showed a marked perinatal increase, and was predominant among system A amino acid transporters. It was first detected in E18.5 liver, and found in most hepatocytes after birth. Three alternative first exons were found in SNAT4 gene. Promoter analyses using ∼3 kb fragments corresponding to each first exon (AP1, AP2, AP3) revealed that AP1 and AP2 exhibited strong promoter activity in mouse hepatoblasts with endogenous HNF4α. Transactivation of AP2 was up-regulated by HNF4α in HeLa cells without endogenous HNF4α. EMSA has demonstrated that HNF4α directly binds to cis-elements in AP2. Over-expression of SNAT4 facilitated amino acid uptake and de novo protein synthesis in primary hepatoblasts. CONCLUSION: SNAT4 functions downstream of HNF4α, and plays significant roles in liver development through mechanisms of amino acid uptake and protein synthesis.
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