title: Transcriptome profiling identifies p53 as a key player during calreticulin deficiency: Implications in lipid accumulation.
abstract: Calreticulin (CRT) is an endoplasmic reticulum (ER) resident calcium binding protein that is involved in several cellular activities. Transcriptome analyses in CRT knockdown HepG2 cells revealed 253 altered unique genes and subsequent in silico protein-protein interaction network and MCODE clustering identified 34 significant clusters, of which p53 occupied the central hub node in the highest node-rich cluster. Toward validation, we show that CRT knockdown leads to inhibition of p53 protein levels. Both, CRT and p53 siRNA promote hepatic lipid accumulation and this was accompanied by elevated SREBP-1c and FAS levels. p53 was identified to bind at -219 bp on the SREBP-1c promoter and in the presence of CRT siRNA, there was decreased occupancy of p53 on this binding element. This was associated with increased SREBP-1c promoter activity and both, mutation in this binding site or p53 over-expression antagonised the effects of CRT knockdown. We, therefore, identify a negatively regulating p53 binding site on the SREBP-1c promoter that is critical during hepatic lipid accumulation. These results were validated in mouse primary hepatocytes and toward a physiological relevance, we report that while the levels of CRT and p53 are reduced in the fatty livers of diabetic db/db mice, SREBP-1c levels are significantly elevated. Our results suggest that decreased CRT levels might be involved in the development of a fatty liver by preventing p53 occupancy on the SREBP-1c promoter and thereby facilitating SREBP-1c up-regulation and consequently, lipid accumulation.
authors.affiliation: a CSIR-Institute of Genomics and Integrative Biology ; Delhi , India.
authors.forename: Saurabh
authors.initials: S
authors.lastname: Vig
authors.forename: Puneet
authors.initials: P
authors.lastname: Talwar
authors.forename: Kirandeep
authors.initials: K
authors.lastname: Kaur
authors.forename: Rohit
authors.initials: R
authors.lastname: Srivastava
authors.forename: Arvind K
authors.initials: AK
authors.lastname: Srivastava
authors.forename: Malabika
authors.initials: M
authors.lastname: Datta
mesh_terms: D000818:Animals; D001665:Binding Sites; D037282:Calreticulin / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D016000:Cluster Analysis; D015536:Down-Regulation; D020869:Gene Expression Profiling; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism; D006801:Humans; D050356:Lipid Metabolism / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008820:Mice, Obese; D011401:Promoter Regions, Genetic; D060066:Protein Interaction Maps; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D051780:Sterol Regulatory Element Binding Protein 1 / Q000235:genetics / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation; D019014:fas Receptor / Q000378:metabolism
chemical_list: D037282:Calreticulin; C491901:FAS protein, human; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D051780:Sterol Regulatory Element Binding Protein 1; D016159:Tumor Suppressor Protein p53; D019014:fas Receptor
keywords: SREBP-1c; calreticulin; fatty liver; lipid; p53