Tissue expression analysis, cloning and characterization of the 5′-regulatory region of the bovine FABP3 gene

Li, Anning; Wu, Lijuan; Wang, Xiaoyu; Xin, Ya; Liu, Zan

doi:10.1007/s11033-016-4026-7

Cited by 16 publications

(10 citation statements)

References 40 publications

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“…RACE ready 5′-end first strand cDNA was synthesized through SMARTer RACE 5′ Kit (Takara, Beijing, China) PCR was performed for the amplification of RACE ready cDNA using two different gene specific reverse primers (Table S2) located between exon 1 and 2 of the TORC2 gene and Universal Primer A Mix (Takara, Beijing, China) was used as forward primer. Nested PCR was conducted with the conditions as mentioned previously [79,80] using SeqAmp DNA polymerase PCR enzyme (Clontech Inc., CA, USA). Touchdown PCR was performed with the conditions of five cycles at 94 °C for 30 s, 72 °C for 3 min; then, five cycles at 94 °C for 30 s, 70 °C for 30 s, 72 °C for 3 min, and; 25 cycles at 94 °C for 30 s, 68 °C for 30 s and 72 °C for 3 min.…”

Section: Methodsmentioning

confidence: 99%

Function and Transcriptional Regulation of Bovine TORC2 Gene in Adipocytes: Roles of C/EBPγ, XBP1, INSM1 and ZNF263

Khan

Raza

Junjvlieke

et al. 2019

IJMS

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The TORC2 gene is a member of the transducer of the regulated cyclic adenosine monophosphate (cAMP) response element binding protein gene family, which plays a key role in metabolism and adipogenesis. In the present study, we confirmed the role of TORC2 in bovine preadipocyte proliferation through cell cycle staining flow cytometry, cell counting assay, 5-ethynyl-2′-deoxyuridine staining (EdU), and mRNA and protein expression analysis of proliferation-related marker genes. In addition, Oil red O staining analysis, immunofluorescence of adiponectin, mRNA and protein level expression of lipid related marker genes confirmed the role of TORC2 in the regulation of bovine adipocyte differentiation. Furthermore, the transcription start site and sub-cellular localization of the TORC2 gene was identified in bovine adipocytes. To investigate the underlying regulatory mechanism of the bovine TORC2, we cloned a 1990 bp of the 5’ untranslated region (5′UTR) promoter region into a luciferase reporter vector and seven vector fragments were constructed through serial deletion of the 5′UTR flanking region. The core promoter region of the TORC2 gene was identified at location −314 to −69 bp upstream of the transcription start site. Based on the results of the transcriptional activities of the promoter vector fragments, luciferase activities of mutated fragments and siRNAs interference, four transcription factors (CCAAT/enhancer-binding protein C/BEPγ, X-box binding protein 1 XBP1, Insulinoma-associated 1 INSM1, and Zinc finger protein 263 ZNF263) were identified as the transcriptional regulators of TORC2 gene. These findings were further confirmed through Electrophoretic Mobility Shift Assay (EMSA) within nuclear extracts of bovine adipocytes. Furthermore, we also identified that C/EBPγ, XBP1, INSM1 and ZNF263 regulate TORC2 gene as activators in the promoter region. We can conclude that TORC2 gene is potentially a positive regulator of adipogenesis. These findings will not only provide an insight for the improvement of intramuscular fat in cattle, but will enhance our understanding regarding therapeutic intervention of metabolic syndrome and obesity in public health as well.

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Section: Methodsmentioning

confidence: 99%

Function and Transcriptional Regulation of Bovine TORC2 Gene in Adipocytes: Roles of C/EBPγ, XBP1, INSM1 and ZNF263

Khan

Raza

Junjvlieke

et al. 2019

IJMS

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“…Many studies have shown that the storage and distribution of fat are related to age, sex, hormones, fat synthesis, fat decomposition transport, and other factors [9‐12]. Among these factors, fatty acid‐binding proteins (FABPs) are critical mediators of fat storage and distribution [13‐16] that bind fatty acids and other lipid ligands. Epidermal FABPs (EFABPs) are differentially expressed between human omental and subcutaneous adipose tissue [11], and elevated levels of adipocyte FABPs (AFABPs) have been found in pericardial fat tissue and are associated with cardiac dysfunction in obese people [17].…”

Section: Fabpsa Organisms Lfabp (Fabp1) Ifabp (Fabp2) Hfabp (Fabp3)mentioning

confidence: 99%

Tracing the evolution of fatty acid‐binding proteins (FABPs) in organisms with a heterogeneous fat distribution

Zhang

Ren

et al. 2020

FEBS Open Bio

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The distribution of fat among both invertebrate and vertebrate groups is heterogeneous. Studies have shown that fatty acid‐binding proteins (FABPs), which mainly bind and transport fatty acids, play important roles in the regulation of fat storage and distribution. However, the systematic and genome‐wide investigation of FABP genes in organisms with a heterogeneous fat distribution remains in its infancy. The availability of the complete genomes of Caenorhabditis elegans, Callorhinchus milii, and other organisms with a heterogeneous fat distribution allowed us to systematically investigate the gene structure and phylogeny of FABP genes across a wide range of phyla. In this study, we analyzed the number, structure, chromosomal location, and phylogeny of FABP genes in 18 organisms from C. elegans to Homo sapiens. A total of 12 types of FABP genes were identified in the 18 species, and no single organism exhibited all 12 fatty acid‐binding genes (FABPs). The absence of a specific FABP gene in tissue may be related to the absence of fat storage in the corresponding tissue. The genomic loci of the FABP genes were diverse, and their gene structures varied. The results of the phylogenetic analysis and the observation of conserved gene synthesis of FABP family genes/proteins suggest that all FABP genes may have evolved from a common ancestor through tandem duplication. This study not only lays a strong theoretical foundation for the study of fat deposition in different organisms, but also provides a new perspective regarding metabolic disease prevention and control and the improvement of agricultural product quality.

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“…Thus, the upregulation of LPL and FABP3 due to overexpression of CREB1 suggested that it can also help control exogenous uptake and intracellular channeling of FA. The promoter sequence of FABP3 has a CREB binding site and is highly conserved in cattle, sheep and pigs [ 32 ]. Taken together, these results indicate that CREB may play a role in lipogenesis in goat mammary gland as in non-ruminants.…”

Section: Discussionmentioning

confidence: 99%

cAMP Response Element Binding Protein 1 (CREB1) Promotes Monounsaturated Fatty Acid Synthesis and Triacylglycerol Accumulation in Goat Mammary Epithelial Cells

Yao

Yang

et al. 2020

Animals

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cAMP response element binding protein 1 (CREB1) is a member of the leucine zipper transcription factor family of DNA binding proteins. Although studies in non-ruminants have demonstrated a crucial role of CREB1 in lipid synthesis in liver and adipose tissue, it is unknown if this transcription regulator exerts control of fatty acid synthesis in ruminant mammary cells. To address this question, we first defined the expression dynamics of CREB1 in mammary tissue during lactation. Analysis of CREB1 in mammary tissue revealed higher mRNA abundance in mammary tissue harvested at peak lactation. Overexpression of CREB1 markedly upregulated sterol regulatory element binding transcription factor 1 (SREBP1), fatty acid synthase (FASN), acetyl-coenzyme A carboxylase α (ACACA), elongase of very long chain fatty acids 6 (ELOVL6), lipoprotein lipase (LPL), fatty acid binding protein 3 (FABP3), lipin 1 (LPIN1) and diacylglycerol acyltransferase 1 (DGAT1), but had no effect on glycerol-3-phosphate acyltransferase, mitochondrial (GPAM) or 1-acylglycerol-3-phosphate O-acyltransferase 6 (AGPAT6). In addition, overexpressing CREB1 led to a significant increase in the concentration and desaturation index of C16:1 (palmitoleic acid) and C18:1 (oleic acid), along with increased concentration of triacylglycerol. Taken together, these results highlight an important role of CREB1 in regulating lipid synthesis in goat mammary epithelial cells. Thus, manipulation of CREB1 in vivo might be one approach to improve the quality of goat milk.

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Tissue expression analysis, cloning and characterization of the 5′-regulatory region of the bovine FABP3 gene

Cited by 16 publications

References 40 publications

Function and Transcriptional Regulation of Bovine TORC2 Gene in Adipocytes: Roles of C/EBPγ, XBP1, INSM1 and ZNF263

Function and Transcriptional Regulation of Bovine TORC2 Gene in Adipocytes: Roles of C/EBPγ, XBP1, INSM1 and ZNF263

Tracing the evolution of fatty acid‐binding proteins (FABPs) in organisms with a heterogeneous fat distribution

cAMP Response Element Binding Protein 1 (CREB1) Promotes Monounsaturated Fatty Acid Synthesis and Triacylglycerol Accumulation in Goat Mammary Epithelial Cells

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